Medical Virology Introduction: Hepatitis C virus (HCV) infection is silent and infected patient develop

chronic hepatitis with increased risk of cirrhosis, hepatocellular carcinoma and end
Oral Presentation stage live disease. Current treatment with direct acting antivirals (DAAs) is very
effective in achieving sustained virological response (SVR). However, expected drug
Complete Genome Sequencing of Measles Virus Isolates Obtained during 2009- resistance and suboptimal activity against diverse HCV genotypes are major
2017 from different parts of India drawback. Next generation sequencing (NGS) with more coverage will be effective in
studying genetic variations in both wild type and resistance associated variants (RAV)
Sunil R. Vaidya*1, Sunitha M. Kasibhatla2, 3, Divya R. Bhattad1, Mukund R.
and immune response. HCV genotyping by commercial real time PCR assays escapes
Ramtirthkar4, Mohan M. Kale4, Chandrashekhar G. Raut5 and Urmila Kulkarni-Kale2
subtypic variations within genotype that need to be evaluated by direct sequencing
for unusual subtype that may evade SVR.
ICMR-National Institute of Virology1, Pune,Bioinformatics Centre2, Savitribai Phule
Material and Methods: The serum samples from treatment naïve consecutive
Pune University, HPC-Medical & Bioinformatics Applications Group3, Centre for
chronic hepatitis C patients at AIIMS (North India) and JNIMS, Imphal were obtained
Development of Advanced Computing,Pune, Department of Statistics4, Savitribai
to study genetic variation of HCV. Real time PCR for HCV quantitation and genotyping
Phule Pune University
was carried out. The next generation sequencing and NS5b region Sanger sequencing
ICMR-National Institute for Research in Tribal Health5, Nagpur Road, Jabalpur, India.
of HCV genome was carried out Denovo assembly and phylogenetic analysis of strains
were done using web based bioinformatics software.
Keywords: Measles virus, Indian isolates, complete genome sequences.
Result and conclusion: Out of 90 HCV samples collected at AIIMS, Genotype 1 was
Background: Measles is a highly contagious viral disease of major public health
21 % and genotype 3a was 78% (n=71) whereas among 142 north east sample,
concern. Measles virus (MeV) is enveloped, single stranded negative RNA genome
genotype 1 was 29% (n=42), 3 was 60.5% (n=86), 4 was 1.5% (n=2) and genotype 6
(15894 nucleotides) belong to Pramyxoviridae family (genus Morbillivirus). MeV is
was 8.5% (n=12). HCV complete genome cDNA and library preparation was carried
serologically monotypic virus and genetically divided into 24 genotypes i.e. A, B1-B3,
out for NGS. The sequence data were subjected for filtration and exclusion of human
C1-C2, D1-D11, E, F, G1-G3 and H1-H2. World Health Organization has a target for
sequence by mapping with HG19 sequence. The remaining reads were used for
elimination of measles and rubella control by 2020. Government of India is
denovo genome assembly. Three north-east strains from genotype 1 and 3 were
committed on ‘Measles Elimination’ in a phased manner. Hence, a second dose of
sequenced and analyzed. Phylogenetic analysis of NS5b region of north Indian HCV
measles introduced in year 2010 through Universal Immunization Program. As per
strain have shown 3a, 3b, 3g and 3i subtypes within genotype 3 HCV.
policy, suspected measles cases (fever with skin rash, cough, coryza and
conjunctivitis) was monitored and laboratory confirmed by serological/molecular
tests by the WHO accredited laboratory. Methods: Complete genome sequencing of
Delineating the role of DNA damage response during Rotavirus replication
thirty MeV isolates obtained during 2009-17 from six states of India were performed
using standard RT-PCR and sequencing protocols. All these MeV isolates were grown
Sarkar R1, Chawla-Sarkar M1*
on Vero hSLAM cells. Consensus MeV genomes/genes were subjected to
phylogenetic and bioiformatic analysis.
Division of Virology, National Institute of Cholera and Enteric Diseases1, Kolkata,
Results: Phylogeny with global isolates revealed genotype-based spatio-temporal
India.
clustering. Estimated genome/gene-wide nucleotide substitution rates of Indian MeV
Email:[email protected]
are comparable with global isolates. Selection pressure analysis indicated modest
role of positive selection in MeV evolution corroborating with its serologically
Background: Cellular DNA damage response (DDR) machinery is present in all
monotypic nature. Though selection was found to be operational across all MeV
eukaryotic cells to monitor and repair DNA in response to cellular DNA damage.
genes except L-gene in global isolates, no positively selected sites were observed in
Induction of cellular DDR results in the activation of cell cycle arrest, DNA repair
Indian isolates. The mutation noted in the H-protein of Indian MuVs is not expected
pathway or apoptosis. Several studies have shown that both DNA and RNA viruses
to alter the antigenicity and structural stability.
can activate the cellular DDR pathway and hijack this pathway to facilate their own
replication. This study was conducted to check the activation of DDR following
Conclusions: Analysis of Indian and global MeV isolates indicates that the circulating
Rotavirus infection and find out its role in Rotavirus replication.
strains during 2009-17 in India belong to genotypes D4 and D8, which have limited
Methods: Western blotting was performed to check the expressions of cellular and
divergence with no potential impact on antigenicity and efficacy of the current
viral proteins following Rotavirus infection. Expressions of viral RNAs were quantified
vaccine strains in controlling MeV infection to meet elimination target by 2020.
by qRT-PCR. Cellular DNA damage was measured by comet assay. Localization of
cellular proteins were visualized by immunofluorescence technique. Infective virion
Denovo assembly of Hepatitis C virus full-length genome following next generation
particles formation was measured by plaque assay.
sequencing and genetic variations of strains circulating in north and north-east
Results: Results have shown that Rotavirus infection induces the activation of the
India
ATM branch of DDR pathway. ATM activation was found to be independent of cellular
DNA damagebut dependent on double stranded break sensor MRN complex as
Sonu Kumar1, Renu Yadav1, Yogesh Kumar1, Chandreswar Prasad1, Jyotish Kumar Jha1,
knockdown of MRE11 and RAD50 by lentivirus prevented ATM activation. Activated
Kekungu Puro4, Sachin Kumar3, Anoop Saraya1, Shalimar1, Suraj Nongthombam2,
form of ATM, CHK2 and MRN complex components, MRE11 and RAD50 were found
Baibaswata Nayak1
to localize within the viroplasm. Inhibition of ATM and CHK2 proteins by chemical
inhibitors block viral replication as observed by reduced viral protein synthesis, viral
Department of Gastroenterology1, All India Institute of Medical Sciences, Ansari
RNA synthesis and viral titers, suggesting a crucial role of ATM protein in viral
Nagar, New Delhi , Jawaharlal Nehru Institute of Medical Sciences2, Imphal,
replication.
Department of Biosciences and Bioengineering3, IIT Guwahati,4 ICAR Research
Conclusion: Overall, these results suggest that rotavirus infection induces the
Complex for NEH Region, Meghalaya
activation of ATM branch of cellular DDR and exploit this pathway for its own
replication.
Email: [email protected]
Characterization of Human papillomavirus type 16 major capsid protein variants as were validated using clinical samples. In addition, dry down RT-PSR reagents were
markers of cervical neoplastic status with functional and phylogenetic analysis prepared and successfully evaluated.
The in-house magnetic bead based nucleic acid extraction followed by dry down RT-
Mane A1, Patil L1, Limaye S2, Kulkarni-Kale U2 PSR assay was found to be highly sensitive and specific. The sensitivity was also
compared with commercially available viral RNA extraction kit and real time RT-PCR.
ICMR-National AIDS Research Institute1, Bhosari, Pune, India, Savitribai Pune The detection limit of dry down RT-PSR was found to be lO copies of RNA transcript.
University2, Ganeshkhind Road, Pune, India This novel methodology will help in developing a rapid, field compatible on-site
detection platform as a concept for
Key words: Human papillomavirus 16; L1 variants; cervical intraepithelial neoplasia; viral agents.
epitopes
Background: The etiological role of infection with high-risk Human papillomavirus One-Step Single Tube Accelerated Quantitative Nucleoprotein Gene Specific
(HPV) types and cervical cancer is well established, while HPV type 16 is most Reverse Transcription Loop
prevalent globally and in India. In a given HPV type, the genome is further classified
into ‘variants’ if presence of nucleotide variations is <2% in coding regions. Major Email:[email protected]
capsid or L1 protein is corner stone in HPV structure. L1 variants can affect viral
assembly and restrict immunological recognition. Developing information on L1 Background:Ebola virus (EBOV) causes severe hemorrhagic fever in humans and
variants is imperative for rational design of newer diagnostics and vaccine. The nonhuman primates with high mortality rates. The Centres for Disease Control and
present study was carried out to characterize HPV16 L1 variants by cervical Prevention has classified them as “Category A Bioterrorism Agents” as they could be
intraepithelial neoplastic status, evaluate their impact on epitopes and determine easily transmitted, result in high mortality, cause major public health impact and
their phylogeny. panic, and require special action for public health preparedness. The recent outbreak
of Ebola virus in West Africa highlights the urgent need to develop a rapid and real-
Methodology: HPV16 L1 open reading frame was analyzed in 152 well characterized time detection system for early clinical diagnosis for better patient management.
archival HPV16 positive cervical samples (48 with normal, 51 with low-grade and 53 Isothermal amplification of nucleic acids has emerged as a promising alternative in
with high-grade cervical status). Intratype variants were analyzed using PCR-directed which rapid and efficient amplification is achieved at a constant temperature without
sequencing, impact on experimentally validated epitopes was predicted in silico, the thermal cycling required in PCR. A one-step single tube accelerated quantitative
while phylogenetic analysis was carried out using MEGA software with maximum reverse transcription loop mediated isothermal gene amplification (RTLAMP) assay
likelihood method. was developed by targeting the nucleoprotein (NP) gene of Ebola virus. The RTLAMP
assay was found to be specific for Ebola virus, without having any cross reactivity with
Results: A total of 58 nucleotide polymorphisms were detected, of which 26 were related haemorrhagic fever viral agents. The comparative evaluation of Ebola virus
novel and 20 non-synonymous. Commonest L1 variants were A6432G (73.7%), NP gene specific RTLAMP assay with RTPCR and Taqman real-time RT-PCR
G7058T (17.8%) and C6852T (16.4%). L1 variants C5862T (H102Y), C6163 (T202N), demonstrated that RTLAMP was 10 to 1000 fold more sensitive than Taqman real-
A6178C (N207T), A6693C (T379P) and A6801T (T415S) showed significant association time RT-PCR and conventional RTPCR respectively with a detection limit of 1 copy
with high-grade cervical disease and an increasing trend with cervical disease number. Ebola virus RTLAMP assay for clinical diagnosis was evaluated with different
progression. Predominance of European lineage (84.2%) was observed, while non- kind of spiked body fluids including serum, urine, saliva, semen and stool samples of
European lineages were associated with high grade cervical intraepithelial neoplasia, healthy human volunteers. The Ebola virus RTLAMP assay could correctly picked up
p=0.012. Ten variants were part of experimentally validated B-cell and three of T-cell the spiked samples up to 1 copy of viral RNA without having any matrix interference.
epitopes respectively. Five variants were located in the immuno-genetic FG and HI The monitoring of gene amplification can also be visualized with the naked eye by
capsid loops and could potentially impact anti-HPV vaccine efficacy. using SYBR green I fluorescent dye. Thus, due to easy operation without a
requirement of sophisticated equipment and skilled personnel, the RT-LAMP assay
Conclusion: Results suggest association of specific L1 variants and non-European reported here is a valuable tool as a point of care diagnostic for the rapid and real-
lineages with cervical intraepithelial neoplasia. time detection of Ebola virus in resource limited health care settings of developing
countries.
Development and evaluation of rapid field compatible sample to answer detection
technique for rapid on-site detection of Chikungunya virus. Inactivation of Chikungunya virus by Ultraviolet Light
Shashi Sharma1, Paban Kumar Dash1, M.M.Parida 1Deepak Pardasani2, D.K. Dubey2
Asha maria mathew1, Amol baburao Mun1, Anukumar balakrishnan1
Virology Division1DRDE, Gwalior,Vertox Division2 DRDE, Gwalior,Defence food National Institute of Virology1, Kerala Unit,T.D Medical College Hospital, Vandanam,
research laboratory,Mysore. Alappuzha2, Kerala, India.

Chikungunya virus has re-emerged as an important pathogen causing epidemics in Email:[email protected], [email protected]
several parts of the world since 2004. Predominantly, CHIKV is often associated with
severe persistent arthralgia in small joints. The confirmed circulation of CHIKV in Key words: UV Inactivation, Chikungunya virus, CL-1000 Cross linker.
India, and South-East Asia is a serious concern and cases from all parts of the country Background: Chikungunya virus (CHIKV) is a rapidly emerging arbovirus causing
are reported throughout the year. The rapid molecular detection is required for millions of infections in more than 40 countries. CHIKV is typically a Biosafety level 3
effective control measure against this viral infection, as the virus share similar (BSL-3) pathogen in many countries and handling of CHIKV requires a high standard
symptoms so often misdiagnosed with Dengue and laboratory safety settings. Many studies require the whole virus and the virus to be
Zika virus infection. The current limitation in existing rapid detection assay is the lack handled in Biosafety level-2 setting. A potential solution for managing this problem is
of suitable rapid field compatible sample extraction platform. We report pathogen inactivation without affecting its antigenicity. In the present study, we
standardization of inhouse magnetic bead based nucleic acid extraction method and attempted to inactivate the CHIKV by UV irradiation.
reverse Transcription Polymerase spiral reaction (RT-PSR), an innovative isothermal Material/Methods: Different UV doses were used to inactivate the CHIKV. The
amplification technique for rapid detection of CHIKV. Subsequently both the methods replication status of the inactivated virus was verified in cell lines. Western blot,
electron microscopy (EM) and Immune fluorescence assay were used respectively to diagnostic performance with the HBV-DNA level in CHB patients at tertiary care set
see the antigenicity, structural integrity and entry of the virus into cell lines. up.
Results: The inactivation was complete when a UV dose of 0.09J/cm2 for 3×30s was Methods:A total of 148 CHB virus-infected patients were enrolled in this study.
used and no change in antigenicity and integrity observed. qHBsAg and HBeAg were performed using Chemiluminescent (LIAISON®XL) and
Conclusions: The study concludes that the virus is antigenically stable and could be Enzyme Immunoassay respectively and HBV-DNA were done using Real-time PCR
used in BSL-2 settings for different experiments. (Applied Biosystems®7500). Comparisons among the groups and diagnostic
performance (Receiver operating characteristic; ROC) were analyzed with SPSS v21.
Results:The median qHBsAg of the study population was 4.35 X 103 IU/ml and 45.9%
Immune Modulation of Macrophages during Chikungunya Virus infection; Its were HBeAg seropositive. Out of the total 148 CHB patients, 86.5% were found with
Implications and Future Per detected HBV-DNA (>5 X 102 IU/ml) with the median viral load of 1.91 X 104 IU/ml.
Among the patients with Undetected HBV-DNA (<5 X 102 IU/ml), median qHBsAg was
Email:[email protected] 2.05 X 103 IU/ml. Significant differences were observed in both qHBsAg (p<0.001) and
HBV-DNA (p<0.0001) according to HBeAg sero-positivity. Correlation analysis
Tapas K. Nayak 1, Prabhudutta Mamidi2, P Sanjai Kumar1, Subhransu S. Sahoo1, between qHBsAg and HBV-DNA revealed positive correlation (r= 0.30; p<0.0001). ROC
Chandan Mahish1, Soma Chattopadhyay 2, and Subhasis Chattopadhyay1 curve analysis showed, the area under the curve (AUC) for the qHBsAg level to
distinguish high HBV-DNA (>1 X 105 IU/ml) was 0.65 (p=0.002) highlighting good
School of Biological Sciences, National Institute of Science Education & Research, diagnostic performance. The best cut-off of qHBsAg for predicting high HBV-DNA level
Bhubaneswar1, HBNI, Jatni, Khurda, Odisha,Infectious Disease Biology, Institute of was 2.95 X 103 IU/ml.
Life Sciences, (Autonomous Institute of Department of Biotechnology, Government Conclusion: This study is the first of its kind to quantitate HBsAg in CHB patients from
of India), Nalco Square2, Bhubaneswar, Odisha, India Bangladesh. Use of this marker may bring about an important complementary
evaluation to HBV-DNA among CHB patients during treatment and follow-up.
Background: Re-emergence of Chikungunya virus (CHIKV) infection has referred as a
major public health concern due to its recent worldwide epidemics and lack of Analysis of microRNA expression in Chronic Hepatitis C Virus Infected Patients in
treatment measures. Even though macrophages are known to harbour CHIKV Bangladesh
infection, the concerned regulatory mechanism of the viral replication and the
related immune responses are not well understood. Hossain Sa, Islam SMRb, Jahan Mb, Tabassum Sb, Yasmin Ma1
Materials and Methods: Raw264.7 cells (mouse macrophage cell line) were infected
with CHIKV and viral replications as well as new viral progeny release were assessed University of Dhaka (DU)1 Bangabandhu Sheikh Mujib Medical University (BSMMU),
by flow cytometry and plaque assay, respectively. Furthermore, immune modulation Bangladesh.
of host macrophages and apoptosis were studied through Flow cytometry, Western
blot and ELISA. Key Words:miRNA; HCV; hsa-miR-92; Biomarkers.
Results: Our current observations suggest that expression of CHIKV proteins were Background: MicroRNAs (miRNAs) belong to a class of non-coding RNAs that are
maximum at 8 hpi and the release of new viral progenies were significantly increased secreted by the cells into the circulation. Aberrant circulatory miRNAs profile are
around 12 hpi. Macrophages undergo both extrinsic and intrinsic pathways of associated with different infectious diseases including HCV-related liver diseases. This
apoptosis. The pro-inflammatory mediators, MHCs and B7 were also found to be study has been designed to explore the miRNA profile among chronic HCV patients in
augmented during infection in indicating robust macrophage immune activation. Bangladesh with the goal of finding suitable candidate biomarkers.
Moreover, it was also observed that the MAPKs are activated during CHIKV infection Methods:Expression levels of four miRNAs (hsa-miR-122, hsa-miR-92a, hsa- miR-
and associated with TNF production in the host macrophages in p-c-JUN dependent 200b and hsa-miR-21) in serum samples from thirty-nine patients with HCV viremia
pathway. Further, 17-AAG (a potential HSP90 inhibitor) was found to regulate CHIKV and twenty healthy volunteers were analyzed using Real-time PCR. The relative
infection, apoptosis and pro-inflammatory cytokine/chemokine productions of host expression level of miRNA and diagnostic performance of selected miRNAs were
macrophages significantly. performed with SPSSv21. In addition, miRNA induced target prediction and pathway
Conclusion: Mouse macrophages are analysis were performed using miRSystem and Enrichr web based bioinformatics
tools respectively.
Evaluation and determination of diagnostic performance of quantitative HBsAg in Results: Relative expression analysis revealed that all the miRNAs except hsa-miR-21
Chronic Hepatitis B virus infected patients of Bangladesh were up-regulated (P=0.002 for hsa-miR-122 and P<0.0001 for both hsa-miR-92a and
hsa-miR-200b) in the sera of HCV patients compared with that in healthy controls.
Islam SM1, Jahan M2, Tabassum S3 Furthermore, the receiver operating characteristic curves showed that hsa-miR-122
(Area under curve (AUC)=0.73, P=0.002), hsa-miR-92a (AUC=0.95, P<0.0001), hsa-
S M Rashed Ul, Islam1 Munira Jahan2, Shahina Tabassum3 miR-200b (AUC=0.88, P<0.0001) could distinguish HCV patients with desirable
sensitivity and specificity. In addition, correlation analysis indicated serum hsa-miR-
Assistant Professor1 Associate Professor2 Professor3, Department of 92a expression was positively correlated (r=0.41, P=0.01) with HCV viral load.
Virology,Bangabandhu Sheikh Mujib Medical University (BSMMU),Dhaka, Bangladesh Pathway analysis of putative target genes of hsa-miR-92a indicated the involvement
of TGF-β signal transduction, HIF-1 signaling, ECM-receptor interaction and mTOR
Email: [email protected], [email protected] signaling pathways indicating their role in HCV replication, chronic HCV mediated
fibrogenesis and growth dysregulation of cells.
Keywords: CHB patients, qHBsAg, HBV-DNA, ROC curve. Conclusions: This study revealed that, these miRNAs, hsa-miR-92a, in particular, are
Background: Hepatitis B Virus DNA (HBV-DNA) viral load measurements are aberrantly expressed in chronic HCV patients and have the potential to serve as non-
recommended for diagnosis and monitoring of patients on the treatment of Chronic invasive biomarkers for HCV infection. Further functional analysis highlighted the
Hepatitis B (CHB). In addition, quantitative HBsAg (qHBsAg) estimation adjunct to possible association of host miRNAs targeted gene regulation during the course of
HBV-DNA is vital to assess the state of HBV chronicity and therapeutic prognosis. HCV infection.
Therefore, this study was aimed to estimate the qHBsAg and to compare its
Identification and characterization of potential dengue virus receptors in Aedes of which the positives were Adenovirus - 154, Enterovirus - 11 and HSV - 7.
aegypti Preliminary analysis shows that the major causative agent is Adenovirus (34%), which
when genotyped showed mostly genotype 8. Knowledge from this study will pave
Yadav K, Rana S V, Rajagopal R. way for establishment

Gut Biology Laboratory,Department of Zoology University of Delhi-110007, India

Role of Poly A Tail in Hepatitis E Virus Replication Efficiency
Email id: [email protected]
Bhise NA, Mustak SS, Lole KS*
Keywords: Aedes aegypti; dengue virus; protein protein interaction; yeast two
hybrid. Email:[email protected]
Background: Aedes aegypti is a major insect vector for dengue virus transmission
across the world. Dengue virus symptoms involve mild fever to haemorrhagic Keywords: HEV, poly A tail, renilla luciferase, replication
disorders in humans as well as non-human primates. The number of dengue disease BACKGROUND: Hepatitis E virus (HEV) is an important public-health concern and
incidences have increased dramaticallyover the past decade. Till date, no universal major cause of enterically transmitted hepatitis worldwide. HEV is a positive sense
vaccine is available against all four serotypes, hence controlling the population of RNA genome with 7-methylguanosine cap at 5’-end and a poly (A) tail at 3’-end,
vector is the key strategy for regulating the transmission of dengue virus to humans. lacking efficient cell culture system in spite of several attempts to adapt the virus.
The molecular understanding of relationship of virus and vector is critical to design Present study aims to see the influence of poly A tail on HEV replication.
the vector based strategy for controlling the dengue epidemic. MATERIALS & METHODS: HEV subgenomic replicon plasmid harbouring Renilla
Material/methods:A cDNA library derived from the pool of A. aegypti mosquito was luciferase gene (pSK HEV-2 Rluc) with poly A tail of 16A residues was used as
established using SMART (switching mechanism at 5’ end of RNA Transcript) in the template. Primers were designed to introduce deletions or insertions in poly A tail of
combination of welldeveloped and efficient molecular machinery of Saccharomyces the construct by site directed mutagenesis. Two deletion mutants were developed by
cerevisiae. Yeast two-hybrid (Y2H) assays were performed using DENV-2 Envelope removing 3- and 6-A residues from the (mut13: with 13-A residues and mut10: with
protein as a bait for identifying the potential interacting proteins. Yeast clones 10-A residues) respectively and mutants were produced by adding 3 and 6-A residues
growing on 3DO, 4DO and exhibiting the expression of alpha-gal assay were selected to construct (mut19: with 19-A residues and mut22: with 22-A residues) to the
and sequenced. Sequencing data was searched for similarity on NCBI database. original replicon. Plasmid constructs were used as templates to generate HEV RNA
Results: A number of potential mosquito proteins that interact with DENV-2 proteins replicons by in vitro transcription and transfected into human hepatoma cells (S10-
were identified. Protein like apolipoprotein D, phosphatase were identified. The 3). Replication of HEV was monitored by dual luciferase reporter assay.
identified interacting proteins might involve in different biological activities that may RESULTS: 6-fold reduction in replication of mut13 HEV replicon and nearly complete
assist in replication of virus inside the insect vector. inhibition of replication of mut10 replicon was observed, indicating the requirement
Conclusions: Identification of different interacting proteins of dengue virus in the of at least 13-A residues in poly A tail for successful virus replication. Extension of poly
insect vector host is an crucial aspect for the understanding of its transmission A tail by 3- and 6- A residues in mut19 and mut22 resulted in 2-fold and 3-fold
process which can further be useful in designing vector based prevention strategies. enhancement respectively in virus replication as compared to original replicon,
CONCLUSION: Poly (A) tail length plays a very significant role in the HEV replication
efficiency.
A MULTI-CENTRIC HOSPITAL BASED STUDY ON EPIDEMIOLOGY OF
KERATOCONJUNCTIVITIS IN INDIA – A Primer
Screening for high risk Human Papilloma Virus (HPV) genotyping by multiplex Real
Email:[email protected] Time PCR using TOCE technology in womens with cervical abnormalities: a tertiary
hospital based study from Raipur, Chhattisgarh.
Background: Viral keratoconjunctivitis which is the most commonly observed ocular
conditions in clinical practice. Though self-limiting it is associated with morbidity, loss Negi SS*, Bhargava A, Singh P, Hussain N, Aggarwal S.
of man working hours and productivity. Most of the studies/ surveillance programs
for conjunctivitis are targeted towards one or more pathogens. There is paucity of Department of Microbiology,AIIMS, Raipur, Chhattisgarh, India.
published literature spanning entire viral spectrum causing viral conjunctivitis
especially in India, most of the work carried out on keratoconjunctivitis is limited to Key words: Cervical cancer screening, HPV genotyping, Colposcopy, Cytology.
outbreak investigation and characterization. This study was designed to conduct a Background:Cervical cancer is the fourth most common malignancy in females
nation-wide screening for the most common causes of viral conjunctivitis. For this worldwide. The screening of human papillomavirus (HPV) by molecular tests holds
purpose we had identified 4 centres PGIMER, Chandigarh; TNMC &BYL NAIR promise for reducing cervical cancer incidence and mortality in low resource
HOSPITAL, Mumbai; NEIGHRIMS, Shillong ; RMRC, Port Blair along with our centre countries like India. Accordingly in this study high risk (HR) HPV genotyping was
JIPMER, Puducherry. The most common causative agent for viral keratoconjunctivitis evaluated against cytology and colposcopy for primary cervical screening in the
includes Adenoviruses, Enteroviruses, Herpes Simplex viruses (HSV) in their womens with cervical abnormalities.
descending order of disease causing frequency and Chlamydia. Adenoviruses Methods:A total 185 clinical sample of cervical scrapes obtained from womens
(serotypes 3, 4, 7, 8, 19 & 37) are one of the most common causative agents for attending gynecology department for routine cervical examination were processed
keratoconjunctivitis worldwide with recent outbreaks caused by Adenovirus serotype by multiplex Real-time PCR using TOCE technology to target L1 gene to detect and
8. Enteroviruses (EV71 and Coxsackie A24) which are known to cause CNS infections identify the 14 HR-HPV genotype and compared with cytology and colposcopy
are also associated with haemorrhagic conjunctivitis. Herpes simplex virus, unlike the examination.
other two viruses does not cause outbreaks. It is primarily responsible for herpetic Results:Among 185 samples, detection rate of HPV by RT-PCR was found 36.2% (67
corneal involvement classically presenting as geographical ulcers. Chlamydial ocular cases) versus Colposcopy 21.08% (39 cases) and cytology 22.16% (41 cases)(p<0.01).
involvement presents in form of Trachoma and Inclusion conjunctivitis which at times HPV 16 was found most prevalent with detection in 18 cases (8.6%) followed by HPV
may be indistinguishable from viral and hence was included in the current study. At 39 (7cases, 3.78%), HPV 18 (06 cases, 3.24%), HPV 31(04 cases, 2.16%), 68(03 cases,
the end of first year a total of 452 samples have been collected from all the centres, 1.62%), HPV 59(2 cases, 1.08%), and HPV 52(01 case, 0.54%). 26 cases (14.05%) were
found with more than one HR HPV genotypes. PAP smear examination showed ASCUS informatics approach was applied to map the presence of immunodominant B and T-
in 26 cases (14.05%), LSIL in 08 cases (4.32%), HSIL in 02 cases(1.08%), SCC in five cell epitopes of rotavirus structural and nonstructural proteins. A multi-epitope based
cases (2.70%). vaccine constructs was designed encompassing the predicted B and T-cell epitopes
Conclusions:HPV genotyping with high detection rate over other conventional that were selected based on flexibility and accessibility. The allergenicity and
procedures may be used in routine cervical screening for higher probability of antigenicity were predicted to ensure the safety and immunogenic behavior of final
detection of HR-HPV genotypes to allow rapid and pragmatic subsequent vaccine constructs. The physiochemical parameters such as theoretical pI, molecular
management to prevent morbidity and mortality. HPV genotyping also provided the weight, aliphatic index and GRAVY score was also calculated. Further, multiple-
information of unusual higher prevalence of HPV genotype 39 in comparison to threading modeling followed by molecular dynamics simulation confirms the overall
HPV18. The baseline data of prevalence of HR-HPV may help vaccine manufacturer stability of the final vaccine constructs and codon optimization was performed to
to consider these genotypes before preparing future polyvalent vaccines. improve the expression of recombinant multi-epitope antigens in bacteria. The
findings of this study may implicate in developing effective epitope-based vaccine in
Recombinant adeno-associated virus mediated effective inhibition of human controlling rotavirus infection and diagnosis of rotavirus in epidemiological
metapneumovirus in mammalian cells. surveillance.

Prashant Kumar*1, Unnati Singh1, Mansi Srivastava1 Discovery and Validation of Chikungunya Antivirals Targeting Viral Protease

Amity Institute of Virology & Immunology, Amity University Noida Uttar Pradesh1 Tomar S*, Singh H1, Mudgal R1, Fatma B1, Kumar R1.

Background: Development of universal vaccines against human metapneumovirus Molecular Virology Laboratory, Department of Biotechnology1, Indian Institute of
(HMPV), a leading cause of respiratory infection, has been a challenge due to lack of Technology Roorkee, Uttarakhand, India.
long term memory in the infected/ immunized individual. There is an urgent need for
an alternative prophylactic/ therapeutic agent which may restrict the virus infection E-mail: [email protected]
in human cells. Antisense oligonucleotides have been shown to be effective in down-
regulating different virus replications in various models but lack of suitable delivery Keywords: Chikungunya, Structure, Peptidomimetic, Antiviral
systems and toxicity due to constitutive expression limits their use for therapeutic/ Background:Chikungunya virus (CHIKV), a mosquito born pathogenic virus, causes
prophylactic purposes. We aimed to develop an adeno-associated virus based fever, arthritis and encephalitis in humans. Presently, there is no specific
delivery system for induced expression of HMPV specific shRNA and ribozymes in drug/antiviral available for CHIKV treatment. Upon entry into host cell, ssRNA of
target human cells. We developed chimeric constructs encoding shRNAs and CHIKV is directly translated into nonstructural polyprotein precursor P1234 which is
ribozymes specific for human metapneumovirus genome and studied their efficacy processed by virus specific nsP2pro. This nsP2pro mediated processing is an essential
and expression profiles in HEK293 cells. A recombinant adeno-associated virus function in viral life cycle and thus nsP2pro constitutes an attractive target for
(rAAV5), having a modified genome expressing the chimeric construct, was generated antiviral drug development.
and its capacity to introduce the construct in human cells as an episome or integrated In this study, we have elucidated the 3D structure of CHIKV nsP2pro which revealed
part of human chromosome was studied. We further investigated the ability of rAAV the presence of two subdomains; N-terminal protease subdomain and a C-terminal
infected human cells to restrict the hMPV infection. Out of five shRNAs and five MTase-like subdomain. Active site of the protein consists of a catalytic dyad made up
ribozymes designed to target the hMPV genome, two shRNAs and one ribozyme was of cysteine and histidine and is located at the interface of the two subdomains. Detail
effective in down-regulating the viral proteins in HEK293 cells. The shRNA-ribozyme structural analysis and comparison revealed the presence of a flexible loop gating the
chimeric construct could show the additive effect of shRNA and ribozyme in down- active site. This loop possesses conserved residues Asn547 and Trp549. Furthermore,
regulating the viral proteins. rAAV particles expressing the chimeric construct could by mimicking the intermolecular interaction between substrate peptide and the
infect the human cells and establish the episome in infected cells which could further active site residues of nsP2pro, a series of peptidomimetic compounds were
express the shRNAs and ribozymes in these cells. These shRNAs and ribozymes could identified. The conformational stability of these compounds was assessed by
significantly reduce the hMPV load in rAAV infected cells. We concluded that rAAV molecular docking and MD simulation. We developed a FRET based nsP2 protease
particles could serve as a potent delivery vehicle for therapeutic nucleic acids to activity assay to assess the inhibitory potential of identified compounds. Interestingly,
restrict the propagation of HMPV in human cells. two of the identified peptidomimetic compounds were found to possess inhibitory
effect on the nsP2pro enzymatic activity. Inhibition of CHIKV nsP2pro by the tested
Exploring rotavirus proteins to design B and T-cell multi-epitope vaccine using in compounds Pep-I & Pep-II was observed with IC50 values of 34µM and 42µM,
silico approach respectively. The inhibition kinetic studies revealed the inhibition constant (Ki) to be
33.34±2.53 µM for Pep-I and 45.89±4.38 µM for Pep-II. Additionally, these two
Damayanti Yengkhom1, Arpita Devi1, R. Doley2, Nima D. Namsa1 compounds were also validated by cell culture based plaque reduction assay and
found to be significantly inhibiting CHIKV replication in BHK-21 cells at concentrations
Department of Molecular Biology and Biotechnology, School of Sciences, Tezpur in nM range, which is much lower than their cytotoxic concentrations.
University, Assam, India

E-mail: [email protected] Identification and characterization of differentially expressed host proteins

modulated during Chikungunya virus infection
Background:Rotavirus (RV), a dsRNA virus in the family Reoviridae, is the leading
cause of severe acute gastroenteritis in infants and young children of less than five Mamidi. P1, Nayak.T.K2, Chatterjee S1, De.S1, Datey.A1, Chattopadhyay S2 and
years of age. The genome consists of 11 dsRNA segments which codes for six Chattopadhyay. S1*
structural (VP1, VP2, VP3, VP4, VP6 and VP7) and six non-structural proteins (NSP1,
NSP2, NSP3, NSP4, NSP5 and NSP6). Each viral protein has a specific role in viral Institute of Life Sciences, Bhubaneswar1,School of Biological Sciences, National
infection and pathogenesis. There has been less extensive study of primary T- or B- Institute of Science Education & Research2, Bhubaneswar, India
cell responsiveness to rotavirus infection and the functional role of these cells or their
correlation to long term immune protection. In this study, a comprehensive immuno-
Background: The emergence of Chikungunya virus (CHIKV) as a global pathogen Conclusions: We concluded that promoter polymorphisms of BAX -248G>A and BCL2
emphasizes the need to understand the biology of this virus for the development of -938C>A could be used as a biomarker for the detection of EBV associated NPC in
effective control measures. Earlier, it was reported by us that Indian outbreak strain North-Eastern India.
(DRDE-06) exhibits faster replication than CHIKV prototype strain S 27 in vitro.
Therefore, a comparative study was carried out between S 27 and DRDE-06 for
identifying the host factors that are modulated during CHIKV infection. Mutational and structural Insights into West Nile Virus RNA dependent RNA
Materials and Methods: Microarray analysis was carried out for S 27 and DRDE-06 polymerase for Antiviral Discovery
infected Vero cells to understand the modulation of host genes. Out of the many, few
selected genes were studied further to explore their functional significance in CHIKV Kumar R1, Rani R1, Kuhn R J2, Tomar S1;*
infection.
Results: Analysis of microarray data revealed that stress response, translational Molecular Virology Laboratory1, Department of Biotechnology, Indian Institute of
control and MAPK pathways related genes were significantly modulated during CHIKV Technology Roorkee, Roorkee, Uttarakhand,
infection. Out of which, MAP kinase activated protein kinase 3 (MK3) and its isozyme Markey Center for Structural Biology2, Purdue Institute of Inflammation,
partner, MAP kinase activated protein kinase 2 (MK2), belonging to p38MAPK Immunology and Infectious Disease Department of Biological Sciences, Purdue
pathway were chosen for further analysis. Both, simultaneous gene silencing of University, Hockmeyer Hall, 240 S. Martin Jischke Drive, West Lafayette, Indiana
MK2/MK3 through siRNA as well as CMPD1 (MK2a specific inhibitor) treatment
reduced CHIKV progeny release significantly in vitro with respect to control. E-mail: [email protected]
Furthermore, CMPD1 treatment, in CHIKV infected suckling C57BL/6 mice reduced
both CHIKV RNA expression as well as expression of inflammatory cytokines in the Keywords: Flavivirus, NS5, protein, cumulative intensity, molecular docking
serum and showed 100% survival in comparison to untreated mice. Background:West Nile virus (WNV), a member of the Flaviviridae, is a positive-sense
Conclusion: MK2 and MK3 were identified as novel host factors involved in CHIKV single-stranded RNA virus. The replication of viral-RNA genome in flaviviruses is
infection inside the host system which can be targeted to modulate the disease dependent of viral-RNA polymerase. Mutational analysis and residue swapping have
caused by CHIKV. been done to clarify the role of the Nuclear-Localization-Sequence residues in RNA-
dependent-RNA-polymerase (RdRp) activity. Expression and purification of NS5
mutants of WNV-RdRp active-domain with wild-type were done using prokaryotic-
The Association of BAX-248G>A and BCL2-938C>A Polymorphism with the Risk of expression-system and Hi-Trap-nickel-immobilized-metal-ion-affinity-
Epstein Barr virus associated Nasopharyngeal Carcinoma: Prediction of Biomarkers chromatography. The inactive-RdRp-mutant was made by site-directed-mutagenesis
for the Detection of the Disease in North-Eastern Indian Population to substitute the active site GDD-motif with GAA using the QuikChange-mutagenesis
protocol and confirmed by DNA-sequencing. Expression and purification of inactive-
Chatterjee Koustav 1, De S 2, Sahu SK 1, Das P 1, Chattopadhyay NR 1, Choudhuri T1. RdRp-mutant were also done. RdRp activity assay was assessed by the primer-
extension assay. The reaction product was resolved in denaturing-RNA-PAGE and the
Department of Biotechnology, Visva-Bharati, Santiniketan,1,2 WB, India, Institute of bands were visualized using Typhoon-FLA-900scanner. The intensities of the
Life Science, NALCO Square, Bhubaneswar, Odisha,India, observed bands were quantified using Quantity-one-1D analysis software. Further, to
identify potential flavivirus NS5 inhibitors, in silico screening and molecular docking
Email: @visva-bharati.ac.in. was done and three potential molecules have been identified. The RdRp assay was
done for all five constructs with the optimal concentration of all required reagents,
Keywords: BAX, BCL2, biomarker, nasopharyngeal carcinoma, North-Eastern India. incubation temperature and time. Based on the cumulative fluorescence intensity
Background: Pro-apoptotic BAX and anti-apoptotic BCL2 are two key proteins in the analysis, all WNV-POL mutants are active while comparing to Wild-type (100%), the
BCL2 family which are involved in apoptotic regulation. Single nucleotide RdRp activity is 88.18% of 69-70, 54.32% of R398A, 39.76% of AMC and 14.6% of
polymorphisms (SNP) in the promoter region of BAX and BCL2 are reported to have inactive-mutant. The molecular-docking results showed that all the three compounds
altered expressions in many cancers. Here we investigated the association of BAX- with high binding energy and good affinity make molecular contacts with the Motif F
248G>A and BCL2-938C>A polymorphism with the prognosis of EBV associated of the finger domain.
Nasopharyngeal Carcinoma (NPC) in the North-Eastern Indian population. All results conclude that mutants are active but the activity is lesser as compared to
Methods: To detect the presence of EBV, PCR was done. PCR-RFLP and DNA wild-type. Based on that study, the host factors which interact with RdRp for nuclear
sequencing were performed for the genotyping to detect polymorphism from both localization can be used to design small molecule inhibitors against the key-step in
NPC samples and Healthy Controls, collected from different State Hospitals in North- viral life-cycle. Molecular-docking analysis revealed that heterocyclic compounds
Eastern India. Insilico study was conducted to inspect any alteration of transcription identified in silico may be considered as the starting point for the development of
factors’(TFs) binding due to SNP. We also conducted meta-analysis to study the antiviral inhibitors targeting.
consequence of these SNP with the risk of the overall cancer susceptibility.
Results: we have found the presence of EBV in NPC samples. From the genotype and Poster Presentation
allelic distribution, we observed an increased risk of NPC in BAX -248 GA (P=0.009),
combined GA-AA genotype (P=0.005) and A allele (P=0.003). BCL2-938 AA (P= 0.03), Pre-transplant seroprevalence and posttransplant reactivation of opportunistic
combined CA-AA genotype (P=0.02) and A allele (P=0.004) have shown significant viral infections in solid organ transplant recipients: a single tertiary care centre
association with the increased risk of NPC. From the insilico approach, the BAX experience from North India
promoter polymorphisms have shown the reduced and unstable binding affinity for
p53 and sp1 TFs towards the promoter. An increased binding affinity of STAB1 and Gupta S1, Gupta E1, Mahajan S1, Pamecha V2, Mathur R.P.3
cyclic AMP response element (CRE) has observed due to BCL2 polymorphism. In
meta-analysis study, besides some ethnic model no such significant association have Department of Clinical Virology1,Department of Hepatobiliary Surgery2Department of
found between BAX-248G>A or BCL2-938C>A SNP and the overall cancer Nephrology Institute of Liver and Biliary Sciences3, New Delhi, Indifa.
susceptibility.
Email: [email protected]
Background: Herpes group of viruses’ causes significant morbidity and mortality in Conclusion: Our finding of changes in serum miR-155 and miR-21 in JE patients
solid organ transplant (SOT) recipients. Aim was to determine pretransplant reveals that microRNAs deserve investigations as potential biomarkers in disease
seroprevalence of herpes viruses in liver (LTx) and renal transplant (RTx) recipients activity which would be helpful in specific and accurate diagnosis. Correlation study
and donors and determine post transplant viral reactivation. between miRNAs and JE severity suggested that miR-155 and miR-21 can serve as
Methods: We undertook this retrospective study for 177 patients{140 (LTx) and 37 new component to develop possible therapeutic strategies for JE in near future.
(RTx)} from January’ 16 – March’ 2018. Data regarding pre-transplant donor(D) and
recipient(R) serological profiles for CMV IgG, EBV IgG, HSV1&2 IgG AND VZV IgG and Infographics Vs. Text: A survey on symptom based completion rates of Test Report
post transplant DNA viral loads were collected. Descriptive analysis was done and Forms
frequencies calculated.
Ganesan M1, Sekar S1, Swarna S1 and Sudha M1.
Results: Pre-transplant serological profiles for the donors and recipients are as
follows; {(CMV IgG: 160 (90.4%)D+R+, 11(6.2%)D-R+, 3(1.7%) D+R- and 3(1.7%)D-R-)},
VRDL Department of Microbiology1, Govt. Theni Medical College, Theni.
{EBV IgG: 159 (89.9%)D+R+, 13(7.3%)D-R+, 3(1.7%)D+R- and 2(1.1%)D-R-)}, {(HSV IgG:
78(44%)D+R+, 55(31%)D-R+, 13(7.3%)D+R- and 31(17.7%)D-R-)} and VZV IgG:
120(67.8%)D+R+, 32(18%)D-R+, 21(11.9%)D+R- and 4 (2.3%)D-R.Post-transplant CMV E-mail: [email protected].
DNA was positive in 41(23.1%); [37(90.25%) LTx and 4 (6.75%) RTx recipients. Out of
41, 7(17.7%) were high risk (D+R-g), 28 (68.3%) intermediate risk (D+R+) and for Keywords: Test Request Form; Clinical symptoms; Infographics.
remaining 6 patients serostatus could not be determined. Post transplant EBV DNA
Purpose of the research:Successful diagnosis and treatment provided to patients is
was positive in 2(4.8%) LTx patients and both were high risk (D+R-). Median CMV viral
always associated with symptom based clinical investigations as these symptoms will
load was higher in RTx (3.5 x 10 3) than LTx (5.0 x 102) copies/ml (p value < 0.005).
offer critical clues to the diagnosis. The clinical history part of Test Request Form (TRF)
CMV disease was seen in 26/41(63.4%) while 2 LTx patients had tissue invasive
is often incomplete or even sometimes unattended. There is a need to enhance the
disease. Post-transplant CMV reactivation was highest in first month [24 (58%)],
attention of physician and to allow paramedical personnel to complete the form as
followed by second [10(24.3%)] and third month [7(17.7%)]. Median time of
physicians are mostly engaged in clinical examination. Hence, an attempt to improve
occurrence of CMV was 25 days (IQR: 13.5-48 days).
the completeness of clinical history in TRF with Infographics (TRF-I) was evaluated.
The principal results:In this study, equal chance for filling up of both TRF and TRF-I
Conclusions: Pre-transplant serological work-up and documentation of herpetic viral
was given to clinicians (in ward) and paramedical persons (at sample receiving point).
infections (CMV and EBV DNA viremia) as early as in the 1 st month of post-operative
A total of 87 TRFs were compared for the completion rates of symptoms, of these,
period might alert clinicians and guide in effective patient management. However,
percentage of unattended forms were significantly reduced from 23% (TRF) to 6.2%
more studies are needed to confirm our findings.
(TRF-I). The percentage of filling each of the symptoms was found to be increased and
while using TRF-I, greater proportion with more than one symptom per patient was
Serum micro RNA-155 and microRNA-21 as a potential non-invasive biomarker to recorded. Symptoms such as cough, chills, throat pain and knee pain was noted as
track Japanese Encephalitis virus infection among Acute Encephalitis syndrome >80% which was almost neglected in TRFs.
patients. Major conclusions:If the relative value of each symptom is considered, combinations
of symptoms can provide greater reliability in the diagnosis of viral diseases. In
Baluni M1, Fatima T1, Ghildiyal S1, Singh D1, Kumar P1, Zia A1, Tiwari R1, Reddy H2, general, visual content attracts the attention and motivates to complete the request
Dhole T2. form. Thus, the current infographics based filling up of clinical history showed a
greater improvement and adopting infographics in request form will be better for
Sanjay Gandhi Post Graduate Institute of Medical Sciences1, Lucknow, Uttar Pradesh, future use in diagnostic laboratories, primary care and epidemiological studies.
India.
King George Medical University2, Lucknow, Uttar Pradesh, India. Role of Mitochondrial Changes in Rabies Pathogenesis: A Study on Post-mortem
Human Brain Samples
Corresponding author- [email protected]
Harsha PK1, Mani RS1, Srinivas Bharath MM2, Mahadevan A3
Keywords:Acute encephalitis syndrome, Japanese Encephalitis, miRNA-155, miRNA-
21, biomarker, serum. 1Departments of Neurovirology, 2Neurochemistry and 3Neuropathology, National
Background:Japanese encephalitis virus (JEV) is a neurotropic mosquito-borne Institute of Mental Health & Neurosciences (NIMHANS), Bangalore
Flavivirus, mainly prevalent in Asia. It is the most important causative agent of acute
viral encephalitis in humans. Recently, microRNAs (miRNAs) are discovered as a key Keywords: Rabies virus, pathogenesis, mitochondrial dysfunction
regulator of inflammatory and immune responses in various diseases including Background:India has the highest burden of human rabies accounting for almost a
neurological and viral infections. Thus, this study was proposed to explore the role of third of the 61,000 global human deaths due to rabies. The major hurdle in the
miRNAs in JEV pathogenesis and to check whether changes in cellular miRNA development of effective therapy for rabies is the lack of understanding of the basic
expression due to JE virus infection, can be detected in circulation which would be mechanisms involved in the pathogenesis of the disease. Recent studies on cell lines
helpful in diagnosis and treatment. using laboratory-adapted rabies virus have indicated the possible role of
Methods:miRNAs (miR-155 and miR-21) were analyzed in the serum of JEV infected mitochondrial dysfunction in rabies pathogenesis. This study was therefore
patients using quantitative reverse transcription polymerase chain reaction (RT- undertaken to examine the role of mitochondrial changes in human brain samples
qPCR). PCR data was normalized using both exogenous (cel-miR-39) and endogenous obtained post-mortem from patients who succumbed to rabies.
(hsa-miR-93) controls and relative expression level of genes between groups was Material and Methods:Postmortem brain tissues from ten confirmed cases of human
calculated using 2-ΔCT method. rabies, and age-matched non-infected controls each were sourced from the Human
Results:Expression of miR-21 was found significantly upregulated (p=0.003) in JEV Brain Tissue Repository, Department of Neuropathology, NIMHANS. Brain
infected patients as compared to healthy controls whereas, miR-155 showed a mitochondria were prepared from frontal cortex, hippocampus, cerebellum, and
significant decrease in JE patients when compared to healthy controls (p= p=0.004). temporal regions of each brain. Mitochondrial respiratory enzymes complex assays
A negative correlation was observed between miR-21 (r= -0.31) and miR-155 (r= - (complex I, II, III and IV) were performed using standardized methods. Citrate
0.22) levels and JE severity.
synthase assay was performed and used as an indicator of mitochondrial mass (CCHFV), Ebola Virus (EbV), Middle East Respiratory Syndrome Corona Virus (MERS-
internal control for complex assays. CoV), Nipah Virus (NiV), Zika virus (ZikV) – is collated with respect to India.
Results:The activity of respiratory chain complex I activity and complex IV activity Methodology:Search engines/databases Pro-Quest Central, JSTOR, PubMed, Google
were significantly higher in all the four regions of rabies infected brains compared to Scholar were used to retrieve appropriate articles reporting confirmed cases of the
non-infected controls. However, activity of complex II and III did not reveal a mentioned seven viruses. Only for ChikV this condition was not applied. Articles were
significant difference in brain samples from rabies-infected cases Vs controls. also retrieved with an aim to identify the first confirmed report of the mentioned
Conclusion: Results of this first study on rabies-infected human brain sample suggest viruses.
a possible role of mitochondrial dysfunction in rabies pathogenesis. The findings of Results: Except for MERS-CoV, all the studied viruses have at least one confirmed case
this study may help in developing novel therapeutic approaches for treatment of in India. Viruses like ChikV, ChpV, CCHFV and NiV displayed multiple epidemics since
rabies. their first report.
Conclusion: Increasing prevalence of the studied viruses evidently suggests that India
Comparison of Influenza A H1N1 Pdm09 activity in central India between 2015 & might be an endemic country with respect to some of these viruses. The reasons
2017 could be many. But the presence of right conditions and vectors for these viruses to
spread visibly indicates that India can no longer ignore them. In this scenario, regular
Nema R K1, Gupta S1, Agarwal A1, Sharma R1, Kudsia A, Nema S, Biswas D monitoring for these viruses becomes mandatory.
Regional Virology Laboratory, All India Institute of Medical Sciences Bhopal1, Saket
Nagar, Bhopal, 462020 India Identification of potential circulatory microRNA as candidate biomarker for liver
cancer in Hepatitis B virus infection
Email: [email protected]
Neeti Nadda1, Sonu Kumar1, Shashi Paul1, Anoop Saraya1, Shalimar1 &Baibaswata
Keywords: Influenza A H1N1 pdm09, Seasonality, Demography, Disease Burden Nayak
Background: Influenza A H1N1 pdm09 strain accounted for the predominant
Influenza activity in our region in 2015 and 2017; while it was relatively dormant in Departments of Gastroenterology and Radio diagnosis,1 All India Institute of Medical
2016. We surmised that emergence of novel viral strains are associated with Sciences, Ansari Nagar, New Delhi, India
increased pathogenicity and transmissibility of the virus and, hence, should reflect in
the disease characteristics of the outbreaks occurring in 2015 and 2017. We, Email: [email protected]
therefore, were interested in comparing the seasonality, demography, disease
burden and clinical characteristics of these two outbreaks. Key word: Hepatitis B virus, HBV, HCC
Methods: Nasopharyngeal/ oropharyngeal swab samples of Influenza-suspects were Introduction: Hepatocellular carcinoma (HCC) is the second leading cause of cancer-
referred to our laboratory from 17 district hospitals and 4 tertiary care teaching related mortality worldwide. Major risk factor for HCC is underlying Hepatitis B virus
hospitals of Madhya Pradesh throughout the study duration. These samples were (HBV) infection as compare to other causes of chronic liver disease. HBV mediated
processed using CDC RT-PCR protocol using CFX96 Connect Thermal Cycler (Bio-Rad). liver cancer occurs due to chronic inflammation, insertional mutagenesis of viral
Results: In 2015 and 2017, 22.18% (431/1934) and 23.24% (639/2749) of Influenza- genome or trans-activation of viral protein. The microRNAs (miRNAs) are small non-
suspects were positive for InfA, of whom InfA H1N1 pdm09 strain accounted for coding RNA released to circulation and aberrantly expressed many diseases including
88.86% (383/431) and 84.19% (538/639) of cases. In contrast, in 2016 only 10.92% viral disease and cancer. These miRNAs can be considered as circulatory biomarker
(113/1034) of Influenza-suspects were positive for InfA H1N1 pdm09 strain. due to ease of detection and high stability in circulation. In HBV induced HCC, the
Comparing the seasonality of outbreaks, we observed that in 2015 peak Influenza potential of these miRNA as early cancer biomarker or marker for cancer progression
activity was in March while in 2017 the same was recorded in September. need to be evaluated.
Predominant age-group affected in 2015 was 21-30 years (20%), while in 2017 Material and Methods: We have screened 84 circulatory miRNAs that are
patients belonging to 0-5 years and 51-60 years were predominantly affected (16.44% differentially expressed in liver cancer using miRNA PCR Array. The miRNAs relevant
and 16.63%). There was a significantly higher proportion of severely ill patients to HBV-induced liver cancer were selected by differential expression in serum of
(category C; guidelines of MoHFW Government of India) in 2017 (72.72% vs. 48.95%) healthy control, treatment naïve chronic hepatitis B (CHB) patient, HBV induced
(P < 0.001). cirrhotic and HCC patients (n=40). Total RNA isolated from serum were
Conclusion: The differences in the seasonality, demographic and clinical severity of polyadenylated, reverse transcribed and amplified by real time PCR. The differential
the outbreaks suggest the likelihood of the emergence of novel viral variants in this expression was analyzed by comparative Ct method after normalization using
region between 2015 and 2017. It would be prudent to verify this possibility through exogenous Cel miR39.
whole genome sequencing of a representative set of samples from the two Result and Discussion: Differential expression of 84 miRNAs in serum of CHB naïve,
outbreaks. cirrhotic and HCC patients were analyzed by normalization with healthy control.
Twelve miRNAs (miR-126, 17, 195, 19b,19a, 20a, 223, 222, 25, 30d, 92a and miR-16)
Exotic human viruses of India. were differential upregulated ((1.12 to 19 fold) and four miRNAs (miR-100, miR-210,
miR-423 and let-7c) were down-regulated (0.4 to 1.18 fold) in HCC as compare to
Poduri C D cirrhotic and CHB patients. Both upregulated (miR-25-3p and miR-92a-3p) and down
regulated (miR-let-7c and miR-100-5p) miRNA were evaluated in validation cohort of
Independent Researcher and Freelance Writer healthy, CHB and HCC patients.
Conclusion: These circulatory miRNAs have potential for cancer marker in HBV
Email: [email protected] infection.

Keywords. ChikV, ZikV, India, Epidemiology Activity of Haemorrhagic Fever viruses in North East Region of India
Background: Infections of a number of exotic human viruses have been reported
from India. In the present study, epidemiology of seven viruses – Chandipura Virus Prasad AK1, Phukan AC2, Barman B3
(ChpV), Chikungunya virus (ChikV), Crimean-Congo Haemorrhagic Fever Virus
Department of Microbiology1,2Department of Medicine3,North eastern Indira Gandhi relatedness. Differences in amino-acid residues encompassing the antigenic epitopes
Regional Institute of Health and Medical Sciences, Shillong India. of VP4 and VP7 capsid proteins between vaccine and Kolkata strains were observed.
Conclusion: GARV shows a tendency towards directional selection under persistent
Key words, Dengue haemorrhagic fever, North East India, Viral Haemorrhagic Fever immunological pressure that might promote natural selection of one genotype over
Backgrounds: Viral Haemorrhagic Fever (VHF) is a major public health problem of another in a particular geographical area. Therefore, surveillance of the circulating
North East (NE) India. The present study was undertaken with the objective to study GARV genotypes shall underscore the importance of introduction of rotavirus vaccine
the etiology of VHF of Dengue, Chikungunya and Crimean Congo Haemorrhagic Fever in this region.
(CCHF) virus origin among patients attending tertiary health care centre in NE India
and also to study the clinical profile of such patients.
Methods: Serum was collected from 51 suspected VHF patients and IgM antibody Changing pattern of Dengue infection in the state of Odisha, Eastern India
capture ELISA was employed to detect anti-Dengue and anti-Chikungunya IgM
antibody. The samples were also tested by real-time RT-PCR for detection of Dengue, Subhra Subhadra1, Jyotsnamayee Sabat2, Bhagirathi Dwibedi3, Subrata K Palo4
Chikungunya and CCHF specific nucleic acid. The demographic, clinical and laboratory Sanghamitra Pati5
profile of the patient was noted in detail.
Results: Serum samples of 16 of 51 suspected cases were confirmed to be suffering VRDL1,2.3.4, ICMR- Regional Medical Research Centre, Bhubaneswar, Odisha
from VHF. Among the confirmed cases, 12 were diagnosed with Dengue
Introduction:Dengue has emerged as a major public health challenge in terms of both
Haemorrhagic Fever, one was diagnosed with Chikungunya and there were three with
clinical pattern and epidemiology. Odisha reported first dengue epidemic in year
Dengue and Chikungunya virus co-infection. However, CCHFV infection was not
2010 and continued to report cases each year in epidemic form during post monsoon
detected in our study. Male preponderance was seen among the infected patients
period becoming an endemic phenomenon in the state. Present study depicts the
where 20 to 30 year age group was found to be predominantly affected. Among
changing pattern of Dengue infection with reference to its serotypes and genotypes.
confirmed cases fever (100%), headache (93.8%), petechial rashes (68.8%), ascites
(37.5%) and conjunctival suffusion (18.8%) were the major clinical findings. Two
Materials and Methods:The study included 5320 suspected Dengue cases referred
patients were diagnosed with Dengue Shock Syndrome who later succumbed to the
from different health facilities of the state during 2010-2017. Dengue NS1 antigen
illness. Most patients (41.2%) were farmers by occupation from Meghalaya and
and IgM antibody was done through ELISA. Serotyping was done for the samples
nearby Arunachal Pradesh.
positive for NS1 antigen through RT PCR by amplifying a part of core-pre- Membrane
Conclusion: Information generated from this study may be used for further in depth
gene (CprM) followed by sequencing and phylogenetic analysis.
molecular epidemiology with characterization of the circulating strains of the viruses
in the NE region of the country. This study further reiterates the fact that CCHF virus Result:Dengue NS1 antigen was detected in 53.2% of cases and IgM antibody in 17.7%
infection in humans is still probably absent in this region despite recent serological cases. Dengue serotype 2 (DEN-2) was the only serotype detected in 2010 and 2011
evidence of its activity in sheep, goat and bovine population of NE India. whereas all four serotypes, 1,2,3,4 (DEN-1, DEN-3, DEN-4) were detected among
cases during 2012-2017. Mixed serotypes were detected in 15% of cases; DEN-2 being
Phylodynamics of circulating genotypes in a hospital based rotavirus surveillance dominant (76%). Phylogenetic analysis revealed genotype IV of DEN-2 and genotype
study during 2008-2017 III of DEN-1 and 3 are mainly circulating in this region.

Lo Ma, Banerjee Aa, Mullick Sa, Dutta Sa, Chawla-Sarkar Ma1 Conclusion:High prevalence of Dengue was observed with increased trend from
2010-2017 affecting both urban and rural areas. Severity of illness was found to be
Division of Virology, National Institute of Cholera and Enteric Diseases 1, Kolkata, associated with mixed infection, however in majority of the cases, it was mild to
India. moderate indicating the endemic nature of disease in most parts of the state.

Email- [email protected] Adjuvant role of a NKT cell activating glycolipid with anti-rabies vaccine

Background: Group-A human rotaviruses (GARV) are among the leading etiological Nagaraju R1, Venkataswamy MM1
agents responsible for acute infantile gastroenteritis worldwide. In an on-going
hospital based diarrhoeal diseases surveillance in Kolkata, Eastern India (2008-17), 1Department of Neurovirology, NIMHANS, Bengaluru.
GARV was identified as the most common cause of infantile gastroenteritis. The
circulating strains were genotyped and characterized to understand their Email: [email protected]
phylodynamics prior to introduction of vaccine in India.
Materials and methods: Stool samples were screened from children (≤5 years) with Keywords: CD1d,Rabies vaccine, glycolipid
diarrhea, seeking health care facilities at two hospitals in Kolkata. Screening for VP6 Background:Rabies, a zoonotic disease caused by rabies virus, is a major public health
antigen was done by ELISA. GARV positive samples were genotyped by multiplex concern in many parts of the world, particularly Asia. Worldwide, more than 55,000
semi-nested PCR and subsequent nucleotide sequencing of VP7 (G- type) and VP4 (P- human deaths occur per year due to rabies, of which 20,000 are from India. Stray
type) gene segments. dog populations serve as principal reservoirs and transmitters for rabies virus in India.
Results: Among 7814 stool samples, 2895 (37.04%) were found to be GARV positive. The current pre-exposure anti-rabies vaccination for stray dogs involves two priming
Highest proportion of GARV positivity was found in children of 6-12 months age doses followed by annual boosters, which is a daunting task. Newer simple
group. The rate of infection peaked during cooler months of November to February. vaccination approaches are essential for improved control of rabies. In this proof of
G1, G2, G3, G9 and G12 in conjunction with P[4], P[6] and P[8] genotypes were seen concept study, we investigated the role of Natural Killer T (NKT) cell based glycolipid
to co-circulate in the population. Incidence of G1 peaked in 2008-10 and 2013-15. adjuvant in enhancing anti-rabies vaccine immunogenicity in the murine model.
Meanwhile, G9 was seen to be the preponderant strain of 2011-13. A sharp deflection Methodology:C57BL/6 mice (n=4), were immunized subcutaneously with either 3
from G1 to G3 strains was observed during 2016-17. Polymorphic G3 strains were first doses of rabies vaccine alone or one dose of rabies vaccine with glycolipid adjuvant.
reported in Eastern India during this study period. All the strains clustered distantly After three weeks, serum samples were tested for Neutralizing antibody titers (Nab)
from the vaccine strains in the phylogenetic dendrograms, depicting less genetic by the rapid fluorescent focus inhibition test (RFFIT). Antigen specific T- cells
producing cytokines, IL-4 or IFN-γ, were estimated by ELISPOT assay.
Results:Nab titers in mice immunized with a single dose of vaccine with adjuvant suspected of congenital CMV infection, the seroprevalence of CMV IgM has been
were higher (2-fold) than mice with 3 doses of vaccine alone (1.11 IU against 0.53 IU). found to be 12.5%-20% by various authors.
The frequency of T cells secreting cytokines were higher in mice receiving combined
vaccine and adjuvant, IL-4 (2-3 folds) and IFN-γ (1-2 folds), as compared to mice
Material and Methods: The blood samples were received from the infants especially
receiving vaccine alone.
those with severe intrauterine growth retardation or for small for age neonates for
Conclusion:This study suggests that the glycolipid adjuvant can potentially enhance
determining the IgG and IgM antibodies against CMV infection over the last one year
the rabies vaccine specific antibody titers as well as Th1 and Th2 cellular immune
responses in the murine model. and six months (January 2017 to June 2018). The tests were put up by μ-capture ELISA
for CMV IgM and IgG antibodies as per the manufacturer’s instructions for qualitative
Prevalence of hepatitis B and C virus in pregnant females in a tertiary care hospital detection. Interpretation of the results was done on basis of controls provided with
the kits. A sample was said to be positive for IgM or IgG antibodies if the absorbance
Tuli A1, Singla N2, Gill M3, Chander J4 value was more than cut-off value.

Department of Microbiology GMCH-32 Chandigarh1,2,3,4 Results: Eleven infants were positive for CMV IgM antibodies during the time period.
They were suspected with having congenital CMV infection. The case histories of
Email:[email protected] these infants were studied retrospectively and the details were noted. The age of the
children varied from as less as five days to one year.
KEYWORDS: Hepatitis C virus, Hepatitis B virus, Pregnancy, Perinatal transmission,
Prevalence Conclusion: Congenital CMV is more prevalent than we think and the clinicians should
BACKGROUND: Viral hepatitis in pregnant females through vertical transmission be on lookout for the disease as early identification of the congenital or perinatal
contributes to the compromised prognosis in the new-born along with increased risk CMV infection can ensure adequate treatment and follow-up
of maternal complications. Perinatal transmission of hepatitis B occurs if the mother
has had acute infection during late pregnancy, or if she is a chronic HBsAg carrier Association of interleukin-1β (-511 C/T) and interleukin-10 (-1082 A/G) gene
while hepatitis C transmission occurs mostly around time of delivery. This information polymorphism with Japanese encephalitis disease in North Indian population.
about prevalence of viral hepatitis during pregnancy is essential for the health and
programme planners. Thus, the current study aimed at investigating their prevalence Ghildiyal S1, Baluni M1, Fatima T1, Singh D1, Zia A1 Gaur P1, Tiwari R1, Dhole T N1
amongst antenatal women attending the tertiary care hospital.
MATERIAL AND METHODS:This study was a hospital based cross-sectional study that Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow,UP, India1
included 3070 pregnant women who attended the antenatal clinic of the tertiary care
hospital, during the periods of Jan 2017-June 2018. The sera collected from the Email- [email protected]
patients were tested for HBsAg and anti-HCV using ELISA kit. Positive and negative
control serum samples were run alongside test.
Key Words: Japanese encephalitis, IL-1β polymorphism and IL-10 polymorphism

RESULTS: Out of total 3070 pregnant females, 100 were detected positive for HBsAg Background: Japanese encephalitis (JE) is the most common form of viral encephalitis
which accounts for 3.3%, and 40 were detected positive for HCV which accounted for in many parts of Asia. It is caused by Japanese encephalitis virus which belongs to the
1.3 %. Most common age group with HBV and HCV infection was 20–29 years. Most family Flaviviridae. JE is associated with neuroinflammation in central nervous system
of the positive pregnant females belonged to rural background. and disruption of Blood brain barrier (BBB). Pro-inflammatory cytokines and anti-
CONCLUSION: Universal free screening of all pregnant women for HBV and HCV inflammatory cytokines play a vital role in JE as well as other neurotropic viral
should be done to prevent the next generation from being grappled by chronic diseases. This study evaluated the association of single nucleotide polymorphisms of
hepatitis, cirrhosis and hepatocellular carcinoma. Screening will reduce the interleukin (IL)-1β (-511) and IL-10 (-1082 A/G) with JE virus susceptibility in northern
prevalence, risks of infection and its transmission to the new-born. Indian population.
Material/methods: We performed a case–control study using 85 patients and 85
healthy controls. The polymorphism study has been performed by restriction
fragment length polymorphism (RFLP).
Congenital CMV infection: a report from a tertiary care center in Chandigarh
Results: We analyzed the C/T and C/C genotypes of IL-1β were significantly associated
with higher risk of JE infection (p=0.000 and p=0.007 respectively) whereas A/G
Singla N1, Jain S2,Kaur P1, Chander J1.
genotype of IL-10 was associated with reduced risk of JEV infection (p=0.001). Among
severe cases of JE, only CT genotype of IL-1β was has been found to be associated
Department of Microbiology1, Neonatology2 Government Medical College Hospital, with disease severity (p=0.02).
Chandigarh Conclusion: IL-1β (-511C/T) may affect the host susceptibility to Japanese
encephalitis in North Indian population. IL-1β (-511C/T) and IL-10 (1082 A/G)
Email: [email protected] polymorphism may be associated with severity of Japanese encephalitis disease.

Background: Human cytomegalovirus is increasingly being recognized as important Association of TLR8 gene polymorphisms in two different groups with the risk of
causes of congenital infection. The incidence of congenital CMV ranges from 0.5– Cytomegalovirus infection among pregnant women in North Indian population.
3.0% in all live births. Intrauterine transmission of CMV to the child can occur
irrespective of prior maternal exposure. Most of the congenitally infected infants Tiwari R 1,2, Fatima T1, Baluni M1, Ghildiyal S1,2, Srivastava J.K2, Dhole T.N.1
(85%-90%) are asymptomatic at birth but 5% to 15% of them develop sequelae mostly
as sensorineural hearing loss, visual impairment or delay of psychomotor Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow1,Amity
development. Dense population, overcrowding, low literacy, poor sanitation, University, Lucknow2
unhygienic conditions, etc, are among the various reasons predisposing people of
developing countries and even poorer society to be exposed to CMV. In children Email: [email protected]
 n these test system. Very similar results were deduced from the CPE images. So, as t
Keywords: HCMV, Polymorphism, TLR8, Pregnant women he concentration of inhibitor increase the inhibitory effect on CHIKV increases as co
Background: Human cytomegalovirus (HCMV) causes the most common intrauterine mpared to control where there is no inhibitor.Thus, the compound ABr and BCl may
infections, affecting approximately 40% to 100% of pregnant women. Taking into be considered as a good inhibitor target against CHIKV.Further biochemical and stru
account the immune response to HCMV, the Toll-like receptors (TLRs) have been ctural based investigations to identify the molecular target of these compounds is in
reported to play important role. Data from various studies suggested that the ability progress.
of certain individuals to respond properly to TLR ligands may be impaired by single
nucleotide polymorphisms (SNPs) within TLR genes, resulting in an altered Modulation of Herpes Simplex Virus-1 infection by MBZM-N-IBT
susceptibility to, or course of, infectious or inflammatory disease. In TLR8, the
common Met1Val missense polymorphism has been implicated in a number of Gangamma S,1Jheelam Sarkar1, Veeksheetha1 and Krishnaja Saseendran1
diseases such as tuberculosis and progression of HIV infection. However, data on the
relation between HCMV infections and TLR8 polymorphisms are absent in the National Institute of Technology Karnataka Surathkal1, Mangaluru, India
literature. Therefore, the current study was aimed to describe the role of TLR-8
(Met1Val and 129 C/G) gene SNPs in the occurrence of HCMV infection among Email:[email protected]
pregnant women and healthy control subjects in Northern India.
Methods: In this case-control study, blood samples were collected from 400 pregnant Background: Life-long latency is usually established for HSV-1 infection and that leads
women, including 150 women infected with HCMV, and 250 control women without to recurrence of the infections. It is known to come central nervous system infection
HCMV infection. Infection in pregnant women was determined by HCMV DNA leading to serious illness, in the immunocompromised population. It is also known to
detection. Further TLR8 allele-specific polymorphism using PCR-RFLP was performed cause central nervous system infection leading to serious consequences. Emergence
and result was analysed using χ2 test. of drug resistance to Herpes simplex viruses (HSV) and cross resistance across
Results: In the current study, G/G genotypes at TLR-8 Met1Val genotype was nucleoside inhibitors has warranted development of new non-nucleoside antivirals.
significantly associated with risk for HCMV infection (p=0.003) whereas, TLR-8 -129 MBZM-N-IBT (1-[(2-Methyl benzimidazole-1-yl) Methyl]-2-Oxo-Indolin-3-ylidene]
C/G genotype did not show any such association. Similarly, the frequency of TLR-8 Amino] Thiourea (MBZM-N-IBT)), was investigated in this study for its capacity to
Met1Val G allele was higher in case group as compared to control group and is inhibit HSV-1.As molecular docking analysis suggested its inhibitory capacity against
associated with risk of disease progression (p=0.013). HSV-1.
Conclusions: The current study demonstrates that G/G genotypes of TLR 8 met1val Materials &Methods: In order to understand the effect of MBZM-N-IBT in HSV-1
increase the risk of development of HCMV infection therefore we can say that infection, plaque assay and confocal microscopy were carried out. Viral RNA and
possibly pre-dispose pregnant women to HCMV infection, thus increasing the risk of protein levels were assessed by RT-PCR Western blot and FACS analysis.Time of
congenital cytomegaly development. addition experiment was performed to understand the possible stage of HSV-1
infection that is affected by MBZM-N-IBT.
Evaluation of Antiviral Efficacy of Halogen-substituted Flavonoids in Chikungunya Results: We have shown that the HSV-1 infectious viral particle formation was
Virus. abrogated by 99% in presence of MBZM-N-IBT. Viral mRNA was also reduced by 72%
and 84% for UL9 and gC respectively. MBZM-N-IBT also reduced the protein synthesis
Rani R1, PuranikNinad V2, Singh V1, Srivastava P2, Tomar S1 for gC and ICP8 significantly. While mRNA of ICP8 was not significantly affected, its
protein synthesis was reduced by 47%. Further, time of addition experiment revealed
Department of Biotechnology, Indian Institute of Technology Roorkee1, Roorkee, Utt the capacity of MBZM-N-IBT to inhibit HSV-1 at early, middle and late stages of
arakhand, India,Bioprospecting Group, Agharkar Research Institute2, G.G Agharkar r infection. Other than inhibiting ICP8, this was attributed to the probable inhibition of
oad, Pune,Maharashtra multiple targets including ICP27, UL42, UL25, UL15 and gB which are involved in
various stages of HSV-1 infection.
Email: [email protected]
Conclusion: This is a significant observation considering the fact that unlike
Keywords: Chikungunya, Halogen substituted flavonoids, Antivir nucleoside inhibitors (acyclovir), MBZM-N-IBT is found to target the multifunctional
Background: Chikungunya virus (CHIKV) is a positive, single-stranded RNA virus of ~1 viral protein (ICP8). Hence, this can further be validated and optimized for the
1.8kb long.It belongs to the family of Togaviridae and genus Alphavirus that is typica development of a non-nucleoside inhibitor as an alternative HSV-antiviral.
lly transmitted by mosquitoes.It has three genotypes depend upon the geographical
distribution-Asia, West African and East Central South African. CHIKV contains 2 ORF Abrupt change in distribution of Dengue virus serotypes over a period of two
s, one on the 3'end and other on the 5'end.The 3'end ORF producing the structural p consecutive years in central India.
roteins while the 5'end producing four non-structural proteins (nsP1-4).After 2016, C
HIKV causes the major outbreak in India. Yadav AK1, Agarwal A1, Ansari K1, Prakash A2, Biswas D1
Flavonoids possess various biological and pharmacological activities that are being e
xploited to develop antiviral and antibacterial drug molecules.In this study, Flavonoi Regional Virology Laboratory, All India Institute of Medical Sciences, Bhopal1,
d based inhibitors against infectivity and replication of CHIKV has been investigated. Department of Microbiology, Barkatullah University Bhopal, M.P2, India.
Three flavonoid compounds (Compounds-ABr, BCl, and CF) that are halogenated at v
arious positions were used and the inhibition potential of these compounds against c Keywords: Dengue, Serotype Distribution, Serotyping.
linical strain of CHIKV has been investigated. The relative antiviral activities of these f Background: The serotype distribution of Dengue demonstrates significant
lavonoids compounds against CHIKV were determined by observing the Cytopathic E geographical and temporal variation across the globe. The frequency with which this
ffect(CPE) followed by the plaque reduction assay and then by quantitative real-time distribution changes in a particular geographical area is not exactly known and data
PCR.These assay proved to be reliable and rapid for determining inhibitory concentr from central India on the serotype distribution of Dengue and its association with
ation as well as inhibitory effect well with clinically achievable drug levels. Compoun different clinical correlates has not been widely reported so far. This study was
d ABr shows 92%, 86% and 45% inhibition in the 70uM, 50uM, and 30uM concentrat conducted to compare the relative distribution of Dengue virus serotypes between
ion range respectively while in the same concentration range compound BCl shows 9 2016 and 2017 among cases of Dengue reporting to a tertiary care teaching hospital
5%, 88% and 56% inhibition respectively but Compound CF had no antiviral activity i in Central India.
Methods & Materials: Clinically suspected Dengue cases were confirmed Department of Microbiology1 Sanjay Gandhi Post Graduate Institute of Medical
serologically through the performance of NS1 antigen and/ or Dengue IgM capture Sciences, Lucknow.
ELISA. Dengue-confirmed cases were subjected to RT-PCR using genus-specific
primers and thereafter serotyped using serotype-specific primers. Email: [email protected]
Results: In 2016 out of 201 Dengue cases, 73 patients were positive for RT-PCR and
the circulating serotype found were predominantly DENV-1 (39.6%) followed by Background and Aim: Dengue virus infection is an important emerging disease of the
DENV-2 (34%), DENV-3 (22%) & DENV-4 (4.4%). In the subsequent year of 2017, the tropical and sub-tropical regions and is mainly diagnosed by serological detection of
distribution of Dengue serotypes abruptly changed from the previous year and NS1 antigen and IgM anti dengue antibodies. Since ELISA facilities are not easily
predominantly comprised of DENV-3 (43.2%) followed by DENV-4 (32.8%), DENV-1 available at most diagnostic centres, so most of them use various commercially
(15.1%) & DENV-2 (8.9%). Mixed infection with multiple serotypes was observed in available Rapid Diagnostic Tests (RDTs) kits. This study was designed to access the
21.9% (16/73) and 19.2% (42/219) of cases in 2016 and 2017 respectively. Comparing diagnostic accuracy of 4 commercially available and widely used RDTs for
the duration of illness with the infecting serotype, we observed relatively prolonged serodiagnosis of dengue virus infection in Indian laboratories.
viremia beyond 5 days of illness in significantly higher proportion of DENV-2 infected Material & Methods: The study was conducted at Department of Microbiology,
patients (p= 0.001). SGPGIMS, Lucknow and GSVM Medical college, Kanpur to estimate the sensitivity and
Conclusion: The present study reports a sudden change in the distribution of DENV specificity of following RDTs: (1) Dengue cassette (Panbio, Australia), (2) Bioline
serotypes over two subsequent years in Central India, which could possibly be the Dengue Duo (S D Diagnostics, Korea), (3) Dengue day 1 test (J Mitra and Co, India), (4)
reason for the outbreak in 2017 being almost 3 times larger. We also observed Dengucheck duo (Tulip Diagnostics, India) on 72 confirmed dengue serum samples
significantly prolonged viremia in DENV-2 infected patients compared to patients that were positive by dengue RT-PCR, dengue NS1 & IgM ELISA along with 80 serum
infected with other serotypes. samples from nondengue febrile illness patients.
Results: The majority of the RDTs demonstrated good sensitivity and specificity for
Air pollution and Health: Modulation of antiviral gene expression and viral entry detecting NS1 antigen. Detection of anti-dengue IgM antibodies showed low
processes sensitivity ranging from 27.8% to 77.7%. However specificity was generally moderate
(50 % to 100%) and more consistent across the assays.
Gangamma S, Jheelam Sarkar1, Veeksheetha1 and Krishnaja Saseendran1 Interpretation and Conclusion: We conclude that NS1 antigen detection by RDTs is
reliable for diagnosis of early acute dengue fever; however RDTs are unreliable to
National Institute of Technology Karnataka Surathkal, Mangaluru1, India detect IgM antibodies for diagnosing late acute dengue fever and ELISA should be
performed as far as possible for serodiagnosis of acute dengue virus infection.
Key words: Air pollution, respiratory infections, innate immunity, phagocytosis.
Introduction: According to a World health organization (WHO, 2016) report, acute
lower respiratory infections are associated with ambient air pollution exposure and Comparative evaluation of in-house developed and commercial PCR kit for
cause significant mortality among children less than five years of age. Lower diagnosis of human polyomaviruses (BKPyV, JCPyV)
respiratory tract infection is one of the leading causes of death among children in
India (Bassani et al., 2010). Viral infection was identified in 49% of pneumonia cases Seetha Dayakar, Heera R. Pillai, Radhakrishnan R. Nair
in children (Ruuskanen et al., 2011). Increased rates of viral infection have been Laboratory Medicine and Molecular Diagnostics1,Srinivasa Ramanujan Institute for
associated with both gaseous and airborne particulate (Cienciwicki and Jaspers, Basic Sciences1, Rajiv Gandhi Center for Biotechnology,Bio-Innovation Center (BIC),
2007). However, much less is known about the underlying mechanisms that link air KINFRA Film & Video Park, Thiruvananthapuram, Kerala, India.
pollution exposure and increase in susceptibility to respiratory infections
(Gangamma, 2012). Moreover, there is a relative scarcity of studies on air pollution Email:[email protected]
associated respiratory infections in India (Gangamma, 2018).
Materials and methods:Airborne particulate matter (PM-10) were collected from Keywords: BKPyV, JCPyV, in-house, PCR/Real-time PCR, renal transplant,
Indian cities. PM extracts were used for stimulation of A549 and monocytes. After immunocompromised patients.
stimulation, viral related host gene or antiviral gene expression was assessed using Background: The human polyomaviruses BK (BKPyV) and JC (JCPyV) are non-
real time PCR. There is significant evidence that viral infections can stimulate enveloped circular double-stranded DNA viruses, members of the Polyomaviridae
secondary infection by colonized bacteria. Therefore, we also examined the effect of family. BKPyV reactivation is mainly related to a renal nephropathy and JCPyV
PM and a viral mimic (R848) exposure on phagocytosis. reactivation can induce the progressive multifocal leukoencephalopathy.
Results and conclusion:PM treatment up regulated expression of several key host Material/methods: The aim of this study was to investigate and to compare a novel,
genes such as vATPC, vATPD, TMPRSS2, Furin and Annexin V. Endosome acidification rapid multiplex nested PCR and Real-time PCR using samples from patients with
genes such as vATPCandvATPD are essential for fusion and viral entry processes. suspected nephropathy requiring a renal transplant or immunocompromised
Taken together our results indicate that PM exposure may enhance viral particle patients. Specimens were tested for human polyomaviruses and the results were
entry into lung epithelial cells. But further experimental confirmation is necessary to compared with the commercial assay.
understand the mechanisms and biological pathways. Further, our experiments have Results: The newly developed PCR was used to screen EDTA blood samples from 87
shown that, R848 exposure significantly reduces phagocytosis. Our preliminary patients attending Government Medical College, Thiruvananthapuram. The method
experiments have shown that air pollution exposure may modulate host genes was compared with a commercially available kit employing real-time PCR, for its
involved in viral entry and innate immunity pathways. Implication of such sensitivity and specificity. Similar pathogen profiles were obtained using both assays
modulations due to air pollution exposure should be further investigated. for samples positive for any or both viruses. 14 (16 %) samples were positive for BK
PyV, 7(8.0%) samples positive for JC PyV and 3 (3.5%) sample showed co-infection.
Can Rapid Dengue Diagnostic kits be trusted? A comparative study of commercially Based on Cohen’s kappa test, the strength of substantial agreement (0.85) of
available Rapid kits. commercial assay was good compared to in-house assay.
Conclusions: Overall, our results show that both the Commercial and in-house assays
Atul Garg,1 T N Dhole. performed similarly in clinical validation. Commercial assays are far more expensive
in terms of costs associated with reagents and consumables. The in-house developed
PCR assay is fast and reliable and provides results with high specificity and sensitivity.
Benefits of use of this in-house assay outweigh the costs of the perhaps more CV A9 (n = 1),EV 80 (n = 1), EV 83 (n = 1), EV 97 (n = 2) in the pathogens. Total 63
accepted commercial assay. (10.75%) HBoVs were detected by real time PCR which were further sequenced by
VP1. Out of them 9 (1.5%) were detected as co infection with NPEVs. Phylogenetic
Comparison of HiMedia’s Insta Q series with Applied Biosystems Step One Plus analysis showed 0.9 - 5.6% divergence at nucleotide level among HBoVs.
Real-time PCR machine Conclusion: ECV and HBoV were found as the main etiologic agent in children
suspected with AFP. Molecular typing of these viruses is useful for characterizing
Gohil.D.1 emerging serotypes and their epidemiological investigation.

Consultant, Research and Development Virology and Molecular Biology1HiMedia To study the pattern of interleukin-21 production during the clinical course of
Laboratories Pvt. Ltd. Ghatkopar (West), L.B.S. Marg,Mumbai, Maharashtra, India primary and secondary dengue virus infection

Email:[email protected] Malik N1, Devi B2, Oberoi L2

Background: Real-time polymerase chain reaction (PCR) is considered as a gold Department of Microbiology, GMC, Amritsar
standard method for characterization of influenza. HiMedia Laboratories has
developed Insta Q series of real-time PCR detection system (Insta Q48 M Series Email: [email protected]
&Insta Q96). The present study aimed to compare Insta Q series with Applied
Biosystems Step One Plus Real-Time PCR system for influenza virus diagnosis. Keywords: IL-21, Primary and secondary DEN infection, DHF, DSS
Method: A total of 15 cell culture supernatant samples of influenza A (H1N1) pdm 09, Background:Dengue is the most important arthropod borne viral infection and due
influenza A, influenza B viruses and 05 confirmed negative cases of influenza A (H1N1) to lack of any commercialized vaccines or accepted pharmacological treatments for
pdm 09 infections were included in this study. Fifteen (15) healthy volunteer samples DENV disease, patient management becomes the real challenge. Several studies have
were collected and used as controls. Viral RNA extraction and one-step real-time PCR revealed the clinical relevance of pro-inflammatory cytokine production during DENV
was performed in triplicates using reagents approved by Centre for Disease Control infection, the present study was conducted to evaluate the production of Interleukin
(CDC) for diagnosis of influenza A (H1N1) pdm 09 virus across all nodal centres in 21,a key soluble mediator mainly produced by CD4+T cells, during the clinical course
India. Results: The Insta Q series suitably diagnosed and typed influenza A (H1N1) of primary and secondary DENV infections and its potential association with disease
pdm 09 virus, influenza A and influenza B virus. The Insta Q series also had a very pathogenesis.
good sensitivity with a difference of upto 3 cycle threshold (Ct) values earlier as Material and Methods:The study was carried out in the Department of Microbiology
compared to Applied Biosystems for the detection and quantification of influenza of Government Medical College, Amritsar. Clinically suspected patients of dengue
viruses. Conclusion: This preliminary study demonstrates the usefulness of Insta Q fever presenting to the emergency, outpatient and indoor the Guru Nanak Dev
series for diagnosis of influenza infections. Greater detection sensitivity of influenza Hospital were included in our study from 1st January 2017 to 30th June, 2018. 100
virus using Insta Q series can be attributed to superior bottom-based detection, Samples testing positive for NS1 antigen, designated as confirmed dengue fever cases
scanning of individual wells using the flexible robotic arm, LED based excitation were included in this study. To differentiate among primary and secondary infections,
source with advanced fibre optics transmission technology and reliable photoelectric commercial ELISA kits were used for detecting of titres of IgM and IgG antibodies in
detection technology. The Insta Q series is a Truly Open Real-time Platform i.e. patients. All the primary and secondary dengue fever cases were then subjected to
compatible with competitor’s kits and reagents. Overall, the Insta Q machines can be quatitative IL-21 analysis by Human IL-21 ELISA Kit. Approval of Institutional Ethics
used to monitor the dynamics of influenza infections during routine testing and for Committee was taken.
outbreak preparedness of influenza virus in India and globally. Results:Confirmed 100 dengue cases were subjected to serological profiling and were
differentiated as primary(50%) and secondary(50%) dengue cases. On the basis of
Etiologic Involvement of Enterovirus and Human Bocavirus in Acute Flaccid symptomatic profiling as per WHO guidelines, 59% classical Dengue fever cases, 29%
Paralysis Cases in India. were DHF and 12% were DSS. Mean IL-21 levels were significantly raised in the
diseased group which was 716±544 pg/ml as compared to the healthy controls
Sahu C1, Singh D V1, Baluni M1, Ghildiyal S1, Dhole T N1 75.18±32 pg/ml. Mean IL-21 levels were more in the secondary dengue cases
(939±561 pg/ml) as against the primary (492±425 pg/ml).
Affiliations: Department of Microbiology, Sanjay Gandhi Post Graduate Institute of Conclusion:It was logical to conduct this study, since an insight into the
Medical Sciences, Lucknow, UP, India 226014 understanding of disease pathogenesis helps combat the disease in a profound
manner. The vision regarding choosing this research was to look for better treatment
Email:[email protected] module for the disease using Interleukin 21.

Keywords: Acute Flaccid Paralysis; Enterovirus; Human Bocavirus Isolation of a novel bacteriophage lytic against E.faecalis strains.
Background: Acute flaccid paralysis (AFP), characterized by the rapid onset of
asymmetric paralysis, can be caused by a variety of viral infections or coinfections. Bhardwaj S, Mehta M, Sood S , Sharma J. , Singh H
Besides wild-type and revertant vaccine strains of polioviruses, several nonpolio
enteroviruses (NPEV), Echovirus ECV), Human Bocavirus ( HBoV) and Cardiovirus ( CV) Institute of Dental Sciences and Hospital, Panjab University, Chandigarh.
have also been associated with AFP.
Methods: Total 586 stool specimens were collected in one year from children Email- [email protected]
suspected for AFP. Molecular method for targeting untranslated region (UTR) and
VP1 capsid region was used for detection of human enteroviruses (HEV), human boca Background & Objective- Enterococci particularly E.faecalis is now recognised as one
viruses (HBoV) and saffold viruses in direct clinical specimen. of the major causes of nosocomial infection. Bacteriophages have effective
Results: HEV RNA was detected in 103 (17.6%) of 586 stool specimens by real time bactericidal activity and have various advantages over antimicrobial agents. The
RT-PCR targeting the highly conserved UTR region. Out of them, 71 (12.11%) were objective of the study was to isolate a novel bacteriophage against E.faecalis isolates
NPEV, partially sequenced by VP1 which revealed the prevalence of echovirus (ECV) from periodontitis patients.
19 (n = 6), ECV 11 (n = 7), ECV 18 (n = 4), ECV 33 (n = 5), Cardiovirus (CV) A10 (n = 2),
Materials & Methods- Gingival crevicular fluid sample was obtained by paper point seropositive MSM were 96.1%, 5.9%, and 0%, respectively. All HBsAg positives were
from periodontitis patients. 46 isolates were identified as Enterococci [39 were also positive for anti-HBc indicating chronic Hepatitis B infection. The seroprevalence
E.faecalis (84.78%) and 7 were E.faecium (15.21%)]. Bacteriophage against E.faecalis rates of anti-HAV in HIV seronegative MSM was 95.6%, while all were negative for
was isolated from sewage sample. The phage was propagated and identified using HBsAg and anti-HCV. An increasing seropositivity for anti-HAV with advancing age
transmission electron microscope (TEM). was noted. The prevalence of syphilis was 27.4% in HIV seropositive and 10.5% in HIV
Results-TEM microscopy showed that the phage had a hexagonal head (73nm) and a seronegative MSM.
100 nm long tail resembling phages belonging to Siphoviridae family. The isolated Conclusion:Results show that Hepatitis A is endemic among MSM, however there is
phage was capable of infecting a spectrum of Enterococcal clinical isolates. a low prevalence of Hepatitis B and C infection indicating that prophylactic hepatitis
Conclusion-The role of bacteriophages as strong biotechnological and natural B vaccination and safe-sex counseling may help prevent transmission of hepatitis
therapeutic agents for Enterococcus faecalis in chronic periodontitis can be viruses among MSM who may be engaged in high-risk behaviours.
considered. Key- Words Bacteriophage, Enterococci, E.faecalis,
Seroprevalence of HIV/HCV-coinfection among patients at a tertiary care center in
Susceptibility towards NPC requires EBV infection and specific host factors Eastern India

Nabanita Roy Chattopadhyay1, Koustav Chatterjee2, Piyanki Das1, and Tathagata Prasad N1, Singh N2,Shahi S K3
Choudhuri1
Department of Virology1,Department of Microbiology2, IGIMS Patna, Professor and
Department of Biotechnology1, Siksha Vhabana, Visva Bharati, Santiniketan, Bolpur, HOD, Department of Microbiology, IGIMS Patna
India.
Email: [email protected]
Email: [email protected]
Keywords: Hepatitis C, HCV, HIV, Coinfection, Seroprevalence, Eastern India
Background:Nasopharyngeal carcinoma (NPC) is a rare, confusing, and commonly Background: The human immunodeficiency virus (HIV) causes immunosuppression
misdiagnosed variety of head and neck cancers. Known risk factors for this disease
and if the patient is coinfected with Hepatitis C Virus (HCV), the probability of the
include host genetic factors, Epstein-Barr virus (EBV) infection, and environmental
person coinfected with HIV/HCV developing cirrhosis increases manifold (1). Studies
and dietary factors. Though rare worldwide, NPC shows a strong ethnic and on HIV/HCV-coinfected patients is highly recommended as it felicitates better
geographic bias for its significantly higher occurrence in mongoloid populations
counseling, treatment and follow up care of HIV/HCV-coinfected patients. Keeping
including those of North-East India. Therefore, a possible major role of specific host
this in mind we decided to do a study to determine the seroprevalence of HIV/HCV-
genetic factors is indicated for the generation and promotion of this disease. EBV
coinfected patients in a tertiary care center in eastern India.
infection might accelerate the process by its cancer causing properties; whereas
Methods: A retrospective study from June 2017 to June 2018 was carried out in the
other external factors like diet and environmental inhalants, add to the neoplasticity.
virology lab of the tertiary care center of Eastern India in which records of all patients
Keywords: Nasopharyngeal carcinoma, incidence, ethnic variation, undifferentiated
whose sera was screened for detection of Anti HCV antibody and HIV antibody by
NPC, North-East India. BenespheraMicrowell ELISA (IHF) were obtained and analyzed. A total of 37845
samples were screened during the above mentioned period out of which 142 (0.37%
Seroprevalence of viral hepatitis in men who have sex with men from Pune,India ) were found to be positive for HCV antibodies and 166(0.43%) were found to be
positive for HIV antibodies, but none was found to be positive for HIV and HCV
Mane A1, Patil L1, Panchal N1, Yelgate R1, Nair V2, Gangakhedkar R3 coinfection.
Conclusions:Future studies with larger sample sizes are needed to estimate the
ICMR-National AIDS Research Institute1,Pune,Udaan Trust, BudhwarPeth2, seroprevalence of HIV/HCV-coinfection and to understand and address the risk factors
Pune,Indian Council of Medical Research3, Ansari Nagar, New Delhi, India associated with its acquisition.

Email: [email protected] Ribavirin inhibits the Chandipura virus replication in Vero cells

Key words: men who have sex with men; hepatitis virus; HIV infection; Anukumar B1,Mun AB2, Amalmol P3
seroprevalence
Background:Men who have sex with men (MSM) have higher chance of acquiring ICMR-National Institute of Virology 1,2,3 (Kerala unit),Govt.T.D.Medical College Hospital,
viral hepatitis. Furthermore a high prevalence of HIV infection among Indian MSM is Alappuzha, Kerala.
reported. HIV coinfected individuals, exhibit more rapid progression to liver cirrhosis
and hepatic failure. The present study was undertaken to determine the Keywords: Chandipura virus, Ribavirin, antivirals
seroprevalence and risk factors associated with acute viral hepatitis among MSM Introduction: Chandipura virus is an emerging tropical pathogen in India. The virus is
from Pune, India. reported to be associated with encephalitis syndrome in young children. The case
Methods:During 2017-2018, a total of 158 MSM, aged 18-56 years, who self-reported fatality rate is around 55-75% and death ensues within 48 hours of infection. Clinical
having had oral or anal sex with another man in last 12 months were enrolled in management and symptomatic treatment is an only option available to treat the
collaboration with Udaan Trust non-governmental organization, Pune. Blood samples infected patients. Antivirals are preferable for exposed patients. As per the available
were collected for laboratory detection of HBV surface antigen (HBsAg), anti-HBs literatures, no antivirals were reported for treatment of chandipura infection.
antibody, anti-HBc antibody (anti-HBc), anti-HCV antibody, anti-HAV IgG and IgM, Methods: This study aimed to test some antiviral drugs, which are available in the
anti-HEV IgG and anti-HDV IgG. market and approved for human use, against chandipura virus. Seven antiviral drugs
Results:Laboratory results were available for 156/158 samples including, 51 HIV were screened for its effect on Chandipura virus replication in vero cells. Dose and time
seropositive MSM and 105 HIV seronegative MSM. The mean age for HIV-positive kinetics of drugs also studied. Differential expression of viral genes post treatment also
MSM was 29±8 years, while for HIV-negative MSM was 29±8.3 years, with no explored in this study.
significant difference (p=0.341). For HIV seropositive MSM, the mean CD4 count was Results: Out of seven antiviral drugs tested against chandipura virus replication in vero
560.8±349.7 cells/µL. Seroprevalence rates of antiHAV, HBsAg, and anti-HCV in HIV cells, ribavirin showed some promising results. At 50 μg extracellular concentration,
ribavirin inhibited 50% of viral plaque formation in cells. Concentration dependent Result: All the 50 CIN grade 2 or worse samples were found positive for HPV by
inhibition was noticed. More than 90% plaque reduction was observed at higher GP5/GP6 PCR. TruenatTM HR HPV showed a sensitivity of 86% (43/50) for CIN2+,
concentration (150 μg) of ribavirin used in this study. The time dependent plaque specificity found 100% (10/10) and 100% positive predictive value and 58% Negative
reduction was also noticed in ribavirin treated cells. The drug was very effective when predictive value. 68% (34/50) samples showed HPV genotype 16/31 and 18% (9/50)
the cells were treated within an hour of post infection. Similarly, significant inhibition belonged to HPV genotype 18/45.
of plaque observed when the cells treated with ribavirin 4h before infection. Ribavirin Conclusion: Laboratory test have a key role in preventing HPV driven carcinomas and
significantly inhibited the viral gene expression in infected cells. guiding therapeutic interventions. TruenatTM Real-Time Micro PCR system being
Conclusion: This study concludes that ribavirin effectively inhibits the chandipura virus semi-automated technique provide rapid and effective means to identify HPV
replication and is a drug of choice for chandipura virus treatment. However, the genotype.
detailed animal study is required before introducing into treatment for Chandipura
virus infection. Recent Nipah Outbreak and global preparedness: Need of One Health approach

Proportion of Japanese Encephalitis, Dengue and Chikungunya virus infections Kumar S1, Saxena S,
among patients with acute febrile illness in a Tertiary care centre.
Center for Advanced Research (CFAR)-Stem Cell/Cell Culture Unit, King George’s
SarkarA1 *, Majumdar T2, Debnath A , Majumder S4 . Medical University (KGMU), Lucknow1, India,CSIR-Centre for Cellular and Molecular
Biology, Uppal Road, Hyderabad2 India
Email: [email protected]
Abstract: Resurgence of Nipah virus (NiV) in South-East Asia has earned the global
Background: Arboviral diseases are on the rise, especially in the north-eastern region attention in last two decades. Approximately >2 billion of people are currently at risk
of India. After establishment of Viral Research and Diagnosis Laboratory in Tripura, of NiV infection. Recently NiV has struck again in India with at least 19 NiV positive
initiative has been taken to diagnose the arboviruses Japanese Encephalitis (JEV), cases including 17 deaths and 753 suspected cases. Kozhikode, Kerala was the
Dengue (DENV) and Chikungunya (CHIKV) by serology. Aim & Objective: The aim of this epicenter of the outbreak which appeared to be a localized occurrence of NiV. The
study was to determine the proportion of JEV, DENV and CHIKV virus infections among emergence of NiV in India is speculated to be instigated by bats. The treatment is
patients with acute febrile illness in a tertiary care centre with objective 1. To diagnose based only on the supportive care and needs isolation of the patients to reduce
JEV, DENV and CHIKV by Immunoglobulin M capture enzymelinked immunosorbent nosocomial infections. In order to cure, results from the antiviral perspectives are
assay (MAC – ELISA). 2. To determine the clinico – demographic profile of the study suggesting the most efficacious drugs are nucleoside based analogs which targets
group. Materials and Methods: Duration of the study is three and half years (January, NiVRdRp. Most of the candidate vaccines for NiV are in the pre-clinical stages and
2015 to June, 2018). Serum samples were collected from the patients with acute febrile have been tested in various animal models. The rationale of virus transmission from
illness and tested for JEV, DENV and CHIKV. MAC – ELISA was preformed as per the bats to animals or human has not been fully understood. Few studies are available in
protocol of the manufacturer provided by NIV, Pune. Results: Out of 3357 subjects 32% order to anticipate the mode of transmission and risk management strategies to
(1063) were MAC – ELISA positive. Among the viruses JEV predominated with 44% prevent the emergence of NiV in future. Moreover, all the preceding outbreaks of NiV
followed by DENV (31%) and CHIKV (25%). Moreover year wise percentage of positivity in South-East Asia have been reported in between month of December and May. The
shows a steady increase in all the three viruses. JEV and DENV are prevalent in males seasonal pattern of outbreaks might be correlated with the breeding season bats
with 62% and 64% over females i.e. 38% and 36% respectively. However CHIKV is which may results in increased virus shedding and date palm sap harvesting period.
predominated in females with 57%, followed by males (43%). In rural areas JEV Geographically, about two billion people subsist in vicinity inhabited by the Pteropus
positivity percentage is 43%, which is more than DENV (32%) and CHIKV (25%). While bats. Small, containable and isolated outbreaks have always a possibility of large
in urban areas DENV is leading with 47% than JEV (28%) and CHIK (25%). Statistical epidemics in future, as we have learned from the recent outbreaks of Ebola and
Analysis: Graph pad prism 7 data analysis package was used to analyse the data. therefore we need to proceed with a sense of urgency in this regards.
Conclusion: This study highlighted the importance of continuous surveillance for
circulating Arboviral agents with molecular typing to look for the circulating genotypes/ Molecular characterization of the common viral etiological agents causing acute
serotypes effective management and prevention. diarrhea among children up to 5 years in a tertiary care setup of Tripura

Evaluation of the Indian TrueNAT micro Real-Time PCR device for case detection of AnkuritaBhowmik1,Dr. Tapan Majumdar2
High Risk Human Papilloma Virus (HR HPV) for cervical cancer screening in India.
SRF, Rota Viral Project, Agartala Government Medical College (AGMC), Agartala 1,
Singh P*, Negi SS, Bhargava A. Tripura. P.I., Rota Viral Project, Associate Professor2, Department of
Microbiology,Agartala Government Medical College (AGMC), Agartala, West Tripura
Department of Microbiology,AIIMS, Raipur, Chhattisgarh, India.
Introduction:Acute viral gastroenteritis is one of the leading cause of mortality
Key words: Human Papilloma Virus, Cervical cancer, CIN2, Real-Time PCR. among children below 5 years worldwide. In India the mortality rate is 13% per year.
Introduction: High risk Human Papillomavirus particularly HPV type 16 and 18 play a The most notable viral agents causing diarrhoea are Rota virus, Adenovirus,
cardinal role in the etiology of the cervical cancer. Truenat TM HPV-HR, a chip-based Astrovirus, Sapovirus and Norovirus. This study was done to find out the circulating
rapid Real-Time Polymerase chain reaction test requiring less than an hour in DNA viral strains in Tripura.
extraction and amplification of specific E6 & E7 gene for the semiquantitative Aim and Objective:The aim of this study was to determine the role of viruses
detection of high risk HPV type 16,18,31,45 in female cervical specimens was attributing to the cause of diarrhea among children < 5 years of age with the following
evaluated for its diagnostic efficiency. objectives:
Method: 50 cervical intraepithelial neoplasia grade 2 or worse (CIN2) cervical  To estiate the proportion of diarrhoea attributable to viral cause.
specimens along with 10 Negative control samples were processed by TruenatTM  Socio-demographic profile of the study subjects.
HPV HR system targeting E6 & E7 gene, and compared against standard GP5/GP6 PCR  Genotypic characterization of the Rota viral agents.
targeting L1 gene. Methodology:Samples were collected with proper labelling in a sterile container
from cases during the acute illness. All the samples were tested for presence of rota
virus antigen by ELISA (ROTACLONE) as per the manufacturer protocol. All the
samples were stored at -80°C for molecular analysis by PCR. Varicella outbreak in a residential child care institution in North India - Time to
Results:A total of 505 stool samples were collected from January, 2017 to July,2018, revisit Immunization guidelines under the Juvenile Justice Act 2015
out of which 185 (27%) samples were positive for Rotavirus, 0-1 year of age being the
highly prevalent age group and the overall prevalence was higher among the males Aditya Sood , Bhavneet Bharti and Mini P Singh
than females. All though the infection was encountered throughout the year, there
was an elevation in the diarrhoeal cases in the months of December to March. The KEYWORDS: Varicella outbreak, vaccination, Residential child care setting.
most common genotypes of Gr-A rotavirus observed were G3P[8] followed by BACKGROUND : Varicella outbreaks are common in India as varicella vaccine is not
G1P[8], comprising of 62.9% and 31.45% respectively. Some other combinations of included in the Universal Immunization schedule for India. Also, a low seroprevalence
G and P serotypes were also observed such asG2P[8],G1P[4],G9P[8],G8P[10] along of varicella antibodies has been reported among Indian children (Balraj and John,
with some untyped strains.Out of 320 Rota ELISA negative samples, 93 samples were 1994, Lokeshwar et al., 2000, Singh et al., 2015, Singh et al., 2011). The outbreaks are
subjected to molecular screening that showed Adenovirus(21%),Sapovirus common in residential child care settings where children are in close contact with
(6%),Norovirus(5%)andAstrovirus (3%). each other leading to increased transmission among peer groups.
Conclusion:To the best of our knowledge this is the first molecular study of human METHODS:Outbreak occurred in a govt owned residential child care institution with
rotavirus, adenovirus, norovirus, sapovirus and astrovirus in North East India. This 103 children. Case of varicella was defined as a child with a macula-papulovesicular
study calls for a continuous surveillance and comprehensive diarrhoeal disease rash without any apparent cause in children residing in the centre. Swabs collected
control strategy to reduce the burden of diarrhoea among children in Tripura, NE from the easily accessible vesicular lesion in two affected children were found to be
India. positive for VZV-DNA. We line listed the cases and collected detailed clinical
information on age, sex, date of onset, number of lesions (severity), treatment taken,
number of days school missed and out of pocket expenditure. Vaccination of all the
In-silico 3-D structural prediction and characterization of Hepatitis E Virus ‘X- susceptible children in addition to isolation of cases, quarantine of suspects and
Domain’ proper screening for new cases was the major control strategy adopted. However
Thakur V1, Ratho R. K1 necessary sanctions and permissions took time resulting in delay in the outbreak
response. Qualitative data collection included narrative information related to the
Department of Virology, PGIMER, Chandigarh India1 problems faced by caregivers in management of these children.
RESULTS: Out of the 103 children we identified around 30 cases of varicella. The index
E-mail [email protected] case was first identified on 27th January 2018. However it was only in April when the
number of cases peaked that the vaccination as control strategy was initiated. All the
Keywords: Hepatitis E virus, X-domain, 3-D structure, Raptor-X children presented with rash. Severity of the varicella as measured by the number of
Background: The macro domain (X) is found to be ubiquitously present from Bacteria skin lesions < 50 (n=10), 50– 100 (n=5), 100-150 (n=15). No child however required
to humans and in many positive-strand RNA viruses like Rubella, Sindbis and SARS hospitalization. All children missed the school for two to three weeks. The burden of
CoV. So far, HEV ORF1 X domain is known to interact with cellular ADP-ribose protein care was significantly increased for the caretakers and isolation policy was difficult to
(involved in host pathogenesis). However, the detailed physiochemical maintain in the presence of common washroom facility. 83% of the children received
characterization and putative refined structure of HEV X-domain with ligand binding Acyclovir treatment and their mean cost of treatment was Rs 597 ± 228.
active sites is not reported yet. So we proposed in-silico 3-D structure and functional CONCLUSIONS: Residential care settings are emerging as susceptible niche area for
characterization of HEV X-domain which will significantly improve our understanding rapid spread of varicella infection with high cost of medical care. Keeping in mind the
of HEV pathogenesis and replication. burden of care of these children, we propose mandatory inclusion of Varicella
Material and Methods: HEV X-domain sequence was retrieved from NCBI and vaccination under the Juvenile Justice Act 2015 for children in need of care and
characterized by ExPASY server. Crystallization probability was predicted by XtalPred, protection in residential child care settings.
solvent accessibility by Raptor-X and disulphide linkages was predicted by DiANNA
and DISULFIND server. Secondary structure was predicted by PredictProtein, SOPMA, Rapid decline of anti-measles antibodies in a cohort of infants from birth to 12
PROFsec and Raptor-X Property server. 3-D structure was predicted by Phyre2, months of age, calls for an urgent change in vaccination strategy in India.
SwissModel, ITASSER and RaptorX and refined by ModRefiner server. Refined
structure was validated by SAVES, RAMPAGE, QMEAN, Verify3D and ERRAT server. Laxmi Makam, Joseph L. Mathew, Radha Kanta Ratho, Mini P. Singh, Sourabh Dutta,
Finally the model was visualized in PyMol and the active binding site and ligands were Bhavneet Bharti, Vanita Suri, Deepika Rechal Massey.
predicted using RaptorX Binding tool and 3D ligandSite server.
Results: The predicted HEV X domain model was found to be more stable (S2> 0.8), Departments of Pediatrics and Virology, PGIMER Chandigarh.
ordered and compact. HEV X-domain represented high crystallization probability
(score 1), two disulfide bond linkages (Cys16-Cys145 and Cys34-Cys91), higher Introduction:Since 1985, India has been administering a single dose of measles
percentage of alpha helical (34%) and extended strand providing thermodynamically vaccine to all infants at 9 months of age. This age was chosen to balance the
stable nature. RaptorX predicted 3D structure and identified HEV X-domain as disappearance of maternal (transplacental) antibodies with the increasing risk of
putative phosphatase (resemblance with 1spvA). Refined structure with 98.1% developing measles. Measles infection occurs before the age of vaccination in 10-15%
residues in favoured region (Ramachandran plot), verified with Verify3D (85.44% cases, suggesting the absence of immunity well before the age of vaccination. This
residues 3D/1D score ≥ 0.2) server was acceptable predicted HEV X-domain. study was designed to estimate the level of measles specific IgG antibodies in a cohort
Multiplicity of 51 represented a deep binding pocket with 19 binding .residues for of term infants followed from birth to 12 months of age; and the pattern of antibody
three different ligands viz. MES, APR and AR6. decline in them.
Conclusions: Physiochemical properties suggested that the model is stable and in Methodology:We enrolled a cohort of 168 term infants in a prospective longitudinal
ordered form and has higher probability for crystallization which could be tried study and measured serum IgG anti measles antibody levels (AMAL) at birth, 3
experimentally. Three ligands are predicted to bind with active binding site of HEV X- months, 6 months, 9 month (pre-vaccination) and 12 months of age. Maternal serum
domain might prove to be a potential inhibitor to limit the HEV pathogenesis. AMAL was also measured at the time of delivery. Antibody levels >12 U/ml were
considered protective.
Results:The median (IQR) maternal AMAL at delivery was 66.15 (26.23 to 138.80) and
the level in infants at birth was 66.83 (29.04 to 128.79). At birth, 142/168 (84.5%) Email: [email protected]
mothers and 149/168 (88.7%) infants had protective levels. The proportion of
protected infants was 25/90 (27.8%) at 3 months, 2/84 (2.38%) at 6 months and 1/34 Keywords: RSV, Under 5 children
(2.90%) at 9 months (pre-vaccination). Three months after vaccination, 81.2% infants Background:RSV is the most common cause of bronchiolitis and pneumonia in young
had protective levels of antibodies whereas 18.8% did not.Infant protection at birth children in both the community and hospital setting and is associated with high
correlated with maternal protection status (Spearman’s correlation coefficient 0.849, mortality and morbidity in under five children. Ongoing surveillance of the clinical and
p= 0.0001). Similarly, protection at 3 months of age correlated with birth antibody epidemiological parameters of RSV is important for development of preventive and
level (Spearman’s correlation coefficient 0.642, p= 0.0001). management strategies. In view of scarcity of data in central India, the present study
was planned to assess the clinical and epidemiological profile of RSV infection in
Conclusion:Measles IgG antibody levels rapidly decline after birth and reach levels far children <5 years of age in a tertiary care hospital.
below protective level well before nine months of age. The majority of infants are Materials & Methods:This is an ongoing study which included children < 5 years of
susceptible as early as 3 months of age, while all are susceptible by 6 months of age. age who visited our hospital with complaints of lower respiratory tract infection.
This requires consideration of earlier vaccination to protect these infants, which could Nasopharyngeal aspirates were collected and transported to microbiology
have significant public health implications. department in viral transport media. The specimens were processed for RSV reverse-
transcriptase polymerase chain reaction. Severity of respiratory tract infection was
Prevalence of viral hepatitis in multitransfused thalassemic population of Haryana: evaluated by PRESS score in RSV positive cases.
A seven-year retrospective study Results:A total of 30 children were recruited in the study [Till Date]. RSV was detected
in 9 (30 %) cases. 8 RSV positive cases were male and 5 were < 1 year of age.
Lall S1 Gill PS2, Gupta T3, Rathee P, Chaudhary U. Respiratory infections due to RSV were categorised as mild, moderate and severe in
3, 4 and 2 cases respectively. Results till November 2018 will be presented during the
Department of Microbiology,Incharge VRDL1,2,3 lab,PGIMS Rohtak,Haryana; conference.
Department of Microbiology, PGIMS Rohtak Haryana Conclusion:Initial findings of our study demonstrate a prevalence of approximately
30% for RSV infection in under five children with manifestations of lower respiratory
Email:[email protected] tract infection.

Keywords: Thalassemia major, Blood transfusion,Viral hepatitis Seasonality trends of rotavirus associated diarrhea in Indian children <5years of age
Background:Thalassemia is one of the common genetic diseases in the in a multisite hospital based surveillance study; 2012-2016
world.Thoughregular blood transfusion improvesthe overall survival of patients; it
carries a definite risk of acquisition of transfusion transmitted infections. The present Karthick.J, Archana Sriraman, Sidhartha Giri, Gagandeep Kang
study was done to assess the prevalence of viral hepatitis in multitransfused patients
of thalassemia major at our centre. [email protected]
Material/methods:Prospective analysis of retrospectively maintained database of
227patients of thalassemia major receiving transfusion at our institute was done over Department & Institution: Wellcome Trust Research Laboratory, Christian Medical
a period of seven years from Jan 2011-Jan 2018. Demographic details of thepatients College, Vellore
were noted. Results of periodic screening of their sera for hepatitis B surface antigen
(HBsAg) and HCV antibody (HCV Ab) by ELISA were analyzed. Introduction: Rotavirus is the leading cause of severe diarrhea in young children <5
Result:The study sample consisted of 165 boys and 62girls suffering from thalassemia years of age worldwide, and causes approximately 78,000 deaths in Indian children
major and receiving blood transfusion (BT) at our centre. Most of these patients had annually.
received BT from other hospitals also before enrolling at our Centre. The median Aim: To assess the seasonality trends of Group A rotavirus strains in children < 5years
agewastwelve years. Twenty (8.8%) patients (14boys and 6 girls) were HCV Ab of age in a four year multisite hospital based surveillance study from 2012 to 2016.
positive by ELISA and one (0.44%) was HBsAg positive. Out of 20 HCV Ab positive Methods: Stool samples were collected from children <5 years hospitalized with
patients, three were also HBsAg positive by ELISA (coinfection rate 1.32%).The diarrhea from seven sites across India during 2012 to 2016, which included 5 southern
prevalence of acquisition of HCV and HBV infection was directly related to number of (Vellore, Trichy, Kolenchery, Hyderabad, Tirupati) and 2 northern (Delhi, Ludhiana)
blood transfusions received by the patients. The prevalence was significantly less sites. Samples were collected after obtaining informed consent from
compared to blood samples before 2014(p<0.001). parents/guardian. All stool samples were screened for group A rotavirus by ELISA
Conclusion:A large number of HCV infected patients remain undiagnosed clinically (Rotaclone).
due to the absence of symptoms specific to hepatic injury like jaundice. Also use of Results: A total of 5834 samples were collected during the time period of four years
screening tests of blood in blood banks based on antibody detection in infections with (2012-2016) from seven sites. 2069 (35.4%) samples were positive for group A
high incubation periodmay eventually lead to iatrogenic transmission of such rotavirus by ELISA (Rotaclone). The proportion of rotavirus associated diarrhea
infections to patients receiving BT.Thus there is a compelling need to implement the increased during November to February of each year across the northern and
policy of nucleic acid based screening tests in blood banks to reduce transmission of southern sites. However, there was a difference in the proportion of rotavirus
BTrelated viral infections. diarrhea between the two regions during the peak months (range: 41.2%-65.5% in
the north, 31.7%-54.9% in the south).
Conclusion: Although rotavirus associated diarrhea was seen throughout the year,
Clinico-Epidemiological Profile of Respiratory Syncytial Virus (RSV) in children less we found that that the proportion of rotavirus diarrheal episodes was more during
than five years of age: First Report from Central India the colder months of the year (November-February), and this trend was seen in north
as well as south Indian sites.
Nema S1, Nema RK1, Bhatt GC2, Shrivastava V2, Kudsia A1, Raghuwanshi A1,Malik
S2,Biswas D1 RNA-Seq analyses of short- and long-term antiretroviral treated patients with
Kaposi’s sarcoma
Deptt. of Microbiology1,Deptt. Of Paediatrics2, AIIMS Saket Nagar,Bhopal,India
Rajput A, Kumar Archit1 and Kumar M1 differences in the amino acids preferences between effective and non-effective
peptides.
Virology Discovery Unit and Bioinformatics Centre1, Institute of Microbial Technology, Conclusion: This in silico method would be helpful for prediction of viral integrase
Council of Scientific and Industrial Research (CSIR), Sector 39A, Chandigarh, India inhibitng peptides.

Email: [email protected] Prediction and analysis of viral and bacterial MHC class II immunogenic and non-
immunogenic epitopes restricted to human host
Keywords: Kaposi’s Sarcoma–associated herpes virus, Antiretroviral treatment, RNA-
Seq, Differential expression gene analysis Mohd. Shoaib Khan1, Amit Kumar Gupta1 and Manoj Kumar2
Background: Kaposi’s Sarcoma–associated herpes virus (KSHV) is a member of
Herperviridae and known to cause Kaposi’s sarcoma (KS). A recent study by Tso et al Virology Discovery Unit and Bioinformatics Centre, Institute of Microbial Technology,
performed the differential expression gene (DEG) analysis. They utilized DESeq2 Council of Scientific and Industrial Research (CSIR), Sector 39A, Chandigarh, India1
method using hg19 version of human genome to compare KS lesions against normal
tissues among four patients i.e. p22, p23, p32, and p83. However, they haven’t Email: [email protected]
explored the effect of short-term (2 to 3 months, p22 & p23) and long-term (>2 years,
p32 & p83) antiretroviral (ART) treatment. Keywords- Epitopes, MHC-II, Immunogenicity, Vaccine, Virus, Bacteria
Methods: In the present study, we have investigated the DEG pattern among above Background- Identification and designing of better immunogenic epitopes is crucial
two groups of patients. We performed RNA-Seq analysis using tophat2/cufflink for effective vaccine development and remains an important area in the field of
pipeline against latest human reference genome hg38. The DEG profiling was carried immunoinformatics. There are attempts to predict the binding affinity of peptides to
out through cuffdiff method between two categories and mapped on GO processes, MHC class II molecules. However, very few attempts were made to predict the
and KEGG pathways. immunogenecity of the peptides and there is a need and scope for better algorithm
Results: On aligning the transcriptomic data of KS patients with the human and KSHV or organism specific immunogenicity predictor.
genomes (hg38 and NC_009333), we found maximum reads were aligned against Methods- In this, MHC class II immunogenic (positiveP) and non-immunogenic
human genome, with at least 91.20% and 86.70% alignment rate for lesion and (negativeN) epitopes belonging to virusesvir (P7181 and N4675) and bacteriabac
control tissues, respectively. Whereas we observed more reads aligned with KSHV (P2907 and N2329) specific to human host were retrieved from the IEDB v3.10.0.
genome from KS lesion (ranges from 158-1285 reads) as compared to normal tissues Further 15-mer epitope subset datasets were also prepared to develop prediction
(0-36 reads). Further, the DEG analyses (cutoff ±1.5 fold) revealed that most of the models. We have employed Support Vector Machine (SVM) to develop prediction
differentially regulated genes (762Up+1093Down) during the short-term treatment, model using different datasets i.e. vir(P7181+N4675), bac(P2907+N2329), vir-vs-
primarily involved in extracellular matrix and metabolic processes were stabilized bac(P7181+N2907), vir_15mer(P2416+N2409), bac_15mer(P931+N1289), vir-vs-
after long-term treatment. Contrary, because of the long-term treatment, the genes bac_15mer(P2416+N931). These datasets were divided into training/testing (~80%)
involved in cell cytoskeleton become up-regulated and the genes for the hair cycle and validation (~20%) dataset. Training/testing datasets (~80%) were used for the 5-
and keratin filament formation become down-regulated. fold cross validation to develop predictive models employing different sequence
Conclusions: These findings provide the regulation of the genes during long-term ART features viz. amino acid composition (AAC), di-peptide composition (DPC), AAC+DPC
in KS patients, which could assist in understanding the role of host-factors in disease hybrid, binary (BIN) and AAC+DPC+BIN hybrid. Validation dataset (~20%) is used for
biology and may be helpful in diagnostics or therapeutic interventions. the independent performance validation.
Results- During 5-fold cross-validation on training/testing datasets, SVM achieved the
Computational prediction of integrase targeting antiviral peptides maximum accuracy, Matthew`s correlation coefficient (MCC) and Receiver operating
characteristic (ROC) of 67.1%, 0.33, 0.72 for vir(P7181+N4675); 77.58%, 0.55, 0.85
Bhardwaj A, Patel PK, Rajput A, and Kumar M for bac(P2907+N2329); 86.39%, 0.69, 0.93 for vir-vs-bac(P7181+N2907); 68.16%,
Email:[email protected] 0.36, 0.73 for vir_15mer (P2416+N2409); 75.84%, 0.51, 0.82 for
bac_15mer(P931+N1289); 85.84%, 0.66, 0.93 for vir-vs-bac_15mer(P2416+N931)
Background: During the past three decades world has seen many viral diseases respectively. All developed models performed equally well on their respective
outbreaks these include Zika, Ebola, Influenza, Dengue etc. These viral diseases are independent validation datasets.
emerged as a great threat to public health due to shortage of the effective Conclusions- We have developed improved MHC class II epitope prediction method
therapeutics against viruses. Peptides based therapeutics are very promising due to and also model to differentiate virus vs bacterial immmunogenic epitopes specific to
their low toxicity in the host and hence many are in various stages clinical evaluation. human host. We hope this would help in identifying organism specific immunogenic
Antiviral peptides can target various stages of the viral life cycle including integration epitopes as putative vaccine candidate.
of viral genome to host cell. Computational search for antiviral drugs against emerging Nipah virus
Methods: In the present study, we developed a prediction method using 225
experimentally validated peptides targeting integrase from “AVPdb” and “HIPdb” Vinay Randhawa#, Shivalika Pathania#, and Manoj Kumar*
repositories. The prediction models were developed using supervised learning by
support vector machine through five-fold cross validation approach. The overall data Virology Discovery Unit and Bioinformatics Centre, CSIR-Institute of Microbial
set includes 76 highly effective and 127 least or non effective peptides. We used Technology (IMTECH), Chandigarh India.
various peptide features like amino acid composition, dipeptide composition, five
amino acid positions at N and C terminals, and their hybrids. The overall dataset was Email: [email protected]
sub-divided into training/testing T203(76p+127n) and independent validation
V22(14p+8n) data sets for performing internal and external cross-validations. Keywords: Nipah virus, nucleoprotein, virtual screening, antiviral drugs
Results: Best performing hybrid model achieved sensitivity, specificity, accuracy, Background: Nipah virus (NiV) is member of the genus Henipavirus in the family
Matthews’ correlation coefficient and receiver operating characteristics of 63.16 %, Paramyxoviridae, which is considered as an emerging deadly virus due to its
73.23 %, 69.46%, 0.36, 0.3 for training/testing while 78.57 %, 62.50%, 72.73 %, 0.41, pathogenicity, has caused several outbreaks in Asian countries. While no therapeutics
0.74 for independent validation datasets respectively. We have also observed are currently approved against this virus, we aimed to prospect for small molecule
antagonists against NiV.
Methods: We considered NiV-nucleoprotein (NiV-N) as potential target based on its Objectives: The present study was undertaken to describe the various Candida
prime role in transcription and replication of viral mRNA and genomic RNA. NiV-N species causing Candidiasis in HIV patients and its antifungal susceptibility among all
structure was obtained from PDB database (4CO6) and a library of FDA-approved age groups.
drugs from DrugBank. After quality assessment and pre-processing, information of Materials&Methods: A total 26 Candida isolates were obtained from all age groups
interactions between ligands and receptor were obtained by docking-based virtual population in HIV positive samples. These isolates were identified on the basis of
screening. As a post-docking filter, only those ligands that occupy binding pocket and Gram Staining. SDA, CHROM Agar, Germ tube test.
interacting with functionally important residues, inferred on the basis of 3D structure Results: Out of 32 HIV positive patients 26 were positive for Candida infection in
overlap with Respiratory syncytial virus ribonucleoprotein, were considered. which 18 were Candida albicans and 8 were non Candida albicans (NAC). The isolated
Results: After molecular docking and assessment of toxicity risks and drug-relevant Candida species were highly sensitive to amphotericin B, Nystatin, Voriconazole and
properties, several potential candidates for NiV treatment were identified such as resistant to Fluconazole.
Bromocriptine and Paritaprevir, and Simeprevir, which are already reported for their Conclusion: Candida albicans was the most prvalant organism isolated from the HIV
antiviral activity against Zika Virus and Hepatitis C Virus, respectively. Importantly, patients in tertiary care hospital. Variation in susceptibility pattern to antifungal
this analysis also provided some novel candidates against NiV-N. Structure-based agents were seen.
clustering indicated identified molecules to be structurally very distinct, except for
Simeprevir and Paritaprevir that were present in one common group. Differential gene expression pattern of host immune genes in Japanese encephalitis
Conclusion: Our analysis identified FDA approved drugs as potential inhibitors of infected patients
Nipah Virus and we expect these leads to be successful in further experimental
evaluation. Purvita Chowdhury1, Siraj A. Khan1*

Arbovirology Division1, ICMR-Regional Medical Research Centre, NE Region,

Etiology of Viral Hemorrhagic Fever in UP West in Tertiary care hospital Dibrugarh, Assam, India

Teerathanker Mahaveer University in Moradabad Delhi road,TMMC & RC, Email: [email protected]
Moradabad, India.
Background: One of the deadliest viral encephalitis caused by Japanese encephalitis
Background: Viral hemorrhagic fever (VHF) is characterized by severe febrile illness (JE) virus shows clinical manifestations ranging from asymptomatic to severe
along with hemorrhagic manifestations. These infections can progress to high fever, neurological symptoms and even death. The precise pathophysiology for the diverse
multi-organ failure, shock and death in many cases.Many viruses cause viral clinical spectrum of JE has not yet been defined. Studies have postulated that during
hemorrhagic fever but Dengue and Chikunguniya virus are more commonly seen in JE infection, inflammatory cytokines and chemokines are produced after the initial
India however the exact prevalence of these infections is not known in UP West is not recognition of viral antigens through the engagement of TLR pathways. However,
known. there is paucity of knowledge on the expression levels of chemokines and TLRs among
Objectives: this study was aimed to assess the prevalence of Dengue and mild and severely affected JE patients. Therefore, the present study aims to
Chikungunya virus in UP West. understand the expression pattern of chemokines, chemokine receptors and TLRs
Materials&Methods:Cases presenting with acute onset of fever with with two or among mild and severe JE cases.
more of the following symptoms including nausea, vomiting, abdominal pain, Materials/Methods: Gene expression levels of chemokines (CCL2, CCL5), its
dirrhoea, weakness, skin rash, headache, lesions on soft palate, unexplained bleeding respective co-receptors (CCR2, CCR5) and TLRs (TLR 3, TLR7, TLR8 and TLR9) was
from any site or joint pain were included in the study. Serum sample from these cases assessed in peripheral blood mononuclear cells (PBMCs) among mild and severe JE
were tested for Dengue virus IgM antibody, Dengue NS1 Antigen, Chikunguniya virus patients as well as in healthy individuals. RNA extraction and cDNA synthesis was
IgM by ELISA. performed as per the manufacturer’s guidelines. mRNA expression levels were
Result: A total 200 cases were enrolled of which a definite etiology was confirmed in quantified using real-time PCR.
85 cases. Out of the 85 positive cases, 28 cases were reactive for Dengue NS1 Antigen, Results: Our study showed significant down-regulation of chemokines (CCL2, CCL5),
46 were reactive for Chikunguniya virus. Males were more commonly affected as their co-receptors (CCR2, CCR5) and TLR3 in mild JE cases as compared to controls.
compared to females (69% vs 31%). Significant difference of gene expression was observed among mild and severe JE
Conclusion: Dengue virus is the most common cause of VHF in west UP India followed cases for CCL2 (p<0.001), CCL5 (p<0.01) and TLR7 (p<0.05).
by Chikunguniya virus. Both Dengue and Chikunguniya virus are included in NVBDCP Conclusion: In conclusion, our results suggest that the variation in gene expression of
(National vector borne disease control program). Regular surveillance, monitoring chemokine CCL2, CCL5 and TLR7 is associated with JE severity and may be used as a
and preventive measures are required to curtail these diseases thereby reducing the probable marker for JE severe pathogenesis.
mortality and morbidity associated with these diseases.
Hepatitis B core antibody positive liver grafts can be safely given in core antibody
Frequency of Candida infection in HIV patients and its antifungal susceptibility in positive and negative recipients with thorough virological evaluation and
TMMC & RC Moradabad monitoring

Farha, Dr.Shewta R. Sharma, Dr. Umar Farooq Dr. Nitin Kumbhar1, Dr. Reshu Agarwal1, Dr Ashok Choudhury2, Dr Viniyendra
Background: AIDS (Acquired Immunodeficiency Syndrome) is a syndrome caused by Pamecha3, Dr. Ekta Gupta1
a virus called HIV (human Immunodeficiency virus). The disease alters the immune
system, making people more vulnerable to infections and diseases. Candida is an Department of Clinical Virology1; 2 Department of Hepatology; Department of
opportunistic fungal pathogen in patients with HIV. Candidiasis commonly affect the Hepato-Pancreato-Biliary Surgery3, Institute of Liver and Biliary Sciences, New Delhi.
skin and mucus membrane. Its incidence significantly worldwide. Identification of
Candida isolates upto the species level is important. There is increased resistance to Introduction:Hepatitis B core antibody (HBcAb) is a marker of exposure to the virus.
antifungal agents and some species are intrinsically resistant to azoles. The use of HBcAb positive grafts for liver transplantation has potential to expand the
donor pool especially in hepatitis B endemic areas.
 Materials and Methods:In this retrospective study, medical records of consecutive Prevalence of cytomegalovirus infection in patients with inflammatory bowel
living donor liver transplant recipients from January 2015 to August 2018 were disease at a tertiary referral centre: A 12 year Retrospective study
analysed for HBV markers (HBsAg, HBcAb, HBV-DNA) in both donors and recipients
and correlated with post-transplant occurrence of HBV infection in recipients. Gupta M1, Dhole TN2
Results:During the study period, 311 liver transplants were conducted. HBcAb was
found to be positive in 9.6% of the donors (30/311). These 30 HBcAb +ve liver grafts Senior resident, Clinical Virology, Department of Microbiology1Professor and head -
were transplanted to 15 HBcAb positive (4 anti-HBs +ve; 11 anti-HBs -ve) recipients Clinical Virology2Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow
and 15 HBcAb negative (5 anti-HBs +ve, 10 anti-HBs -ve) recipients. In HBcAb +ve
group none of the recipients developed HBV infection during the follow up period Email: [email protected]
(mean 9 months; range 2-22 months). In HBcAb -ve anti-HBs +ve group no recipient
developed HBV infection (mean follow up 10.2 months; range: 2 to 32 months). Keywords: Cytomegalovirus, PCR, Inflammatory bowel disease
However, in the HBcAb -ve anti-HBs –ve group one recipient (1/30; 3.33%) developed Background: Cytomegalovirus (CMV) infection exacerbates IBD refractory to
HBV infection documented at 32 months after transplantation. HBV DNA was 6.8 X immunosuppressive therapies. Outcome of patients with IBD and CMV infection is
107 IU/mL. likely to be worse, if treatment with immunosuppressive drugs is continued without
The sole post-transplant HBV infection was seen in HBcAb negative group and the treating the CMV infection. The current ECCO guidelines recommend colonic tissue
recipient had pre-transplant anti-HBs titer of <10 mIU/mL; below the protective levels PCR as the preferred test for screening for CMV colitis.
recommended. Aim:We examined the diagnostic yield of PCR in suspected CMV infection from
Discussion:HBcAb positive liver grafts can be safely given in core positive recipients intestinal biopsies in IBD patients.
and core negative recipients with protective anti-HBs titres. The pre-transplantation Methods: Retrospective study wherein patients who underwent testing for CMV on
anti-HBs titre in recipients, especially HBcAb negative, is very critical and should be intestinal biopsies were identified from January 2006-March 2018. Patients with
evaluated as anti-HBs defines protection against HBV infection. Careful virological serum CMV PCR or antibody testing were not included.
monitoring is recommended in recipients who demonstrate low levels of anti-HBs Results: From 2006 to 2018, at our institution 55.89% (109/195) of patients with IBD
titres and in non-responders. Further, anti-HBV prophylaxis can be recommended in tested positive for CMV by tissue PCR. of PCR+ patients, majority were on steroids
select cases. either alone or in combination with immunomodulator and/or biologic therapy. CMV
DNA was detected in inflamed colonic tissue 40.81%(20/49), rectal tissue
Identification of potential candidates to design therapeutic vaccine against HPV 16 62.4%(78/125) sigmoid tissue 70%(7/10), rectosigmoid tissue 50%(1/2), ileal tissue
infection: molecular docking study 67%(2/3), duodenal tissue in 33.3%(1/3) and not detected in caecal tissue (0/1). Year
wise distribution of CMV-positive patients and frequency of testing was found to be
Kinikar Ma & Kulkarni-Kale Uaa markedly increasing over the years.
Conclusion: CMV reactivation potentially causes severe colitis in these patients who
Bioinformatics Centre, Savitribai Phule University of Campus, Ganeshkhind Road, are more likely to become hospitalized, have longer lengths of stay and higher
Pune,India mortality rates. Patients with CMV infection are more prone to develop steroid
refractoriness and a cut-off value of CMV DNA to predict the steroid refractoriness is
Email:[email protected] yet to be identified but taken as 250copies/mg. Histology is quite specific but is of low
sensitivity.Therefore, an early detection of CMV DNA in intestinal biopsy is of utmost
Keywords: Cervical cancer; Human papillomavirus; E6; T-cell epitopes, bioinformatics importance for the correct and timely management of IBD patients with CMV
Background: Human papillomavirus (HPV) infection causes genital warts and infection and reduction of colectomy rates.
anogenital, head and neck cancers. It is a sexually transmitted infection found in
almost all sexually active individuals. HPVs are classified as high-risk (cancer inducing) Characterization of E7 gene of HPV16 in Indian population: an evolutionary
and low-risk (cause genital warts). HPV type 16 is a high-risk HPV responsible for bioinformatics study
causing cervical cancer in 70% of women. Prophylactic vaccines are available against
some types of infectious HPVs which have resulted in prevention of HPV infection. Email:[email protected]
But, due to lack of awareness and vaccination, there are cases of women suffering
from cervical cancer due to HPV type 16 infections. Hence, there is a need to develop Background: Human papillomaviruses (HPV) belong to Papillomaviridae family. HPV
therapeutic vaccine against HPV type 16 infections. The HPV 16 genome codes for 8 are non-enveloped, circular DNA viruses with 7.9 kbp genome encoding 7 genes. HPV
proteins (E1, E2, E4, E5, E6, E7, L1, and L2) out of which E6 are E7 are oncoproteins. are classified on the basis of its capsid protein L1 into 5 genera. HPV16, a member to
Epitopes derived from oncogenic proteins could be considered potential targets for Alphapapillomavirus 9, is considered a high risk type known to cause cervical cancer
designing therapeutic vaccine. This study aims to identify such targets from E6 protein in women. E7 gene codes for an oncogenic protein which targets Retinoblastoma
of HPV 16 using docking simulations. tumor suppressor (Rb1) and cause cell proliferation. The study aimed at
Materials/Methods: Experimentally validated T-cell epitopes of E6 were retrieved characterization of variations observed in E7 gene of HPV16 isolates circulating in
from IEDB online (https://www.iedb.org/). These epitopes were docked onto MHC Indian population. The sequencing of 125 isolates was carried out at National AIDS
class I and MHC class II molecules using ZDOCK-RDOCK, Glide (Schrödinger software) Research Institute (NARI). The sequences thus obtained were compared with 50
and ClusPro. Analysis of the docked poses was done using Discovery Studio. reference sequences of global isolates in public domain databases with an objective
Results: Eight epitopes were docked onto HLA*A0201, HLA*B5703, HLA*B2709, to understand extent of evolutionary forces operational in E7. Methodology: The
HLA*DPA1*0103 MHC alleles based on their affinity to a particular allele.The bioinformatics pipeline included multiple sequence alignments, mutation detection,
molecular docking results showed significant interactions between the anchor phylogenetic analysis (alignment-based and alignment-free), mapping known
residues of MHC class I molecules and epitopes. Therefore, epitopes were ranked on antigenic sites and mutations on 3D structure. Results: The nucleotide sequences of
the basis of respective docking scores and experimental validation of CTL responses E7 gene of Indian isolates showed 10 mutations of which 2 are non-synonymous
curated from publications. mutations. Phylogeny of E7 gene helped identify circulating lineages of HPV in Indian
Conclusion: On the basis of the ranking, epitopes ’22-TIHDIILECV-31’ and ’11- population. Majority of Indian isolates belonged to ‘A’ lineage. A total of 5 B-cell and
KLPQLCTEL-19’ are identified as potential targets for therapeutic vaccine design. 15 T-cell experimentally validated epitope of E7 were obtained from IEDB which were
mapped onto the E7 protein structure. Conclusion: E7 gene undergoes a few
mutations and depicts a clustering pattern similar to the lineages obtained using L1 also to study the demographic and clinical profile of such patients.
gene, which is used as a marker for HPV classification. With no recombination and in
Methods: Cerebrospinal fluid(CSF) and serum samples were collected from 357
the absence of positive and pervasive selection E7 gene is highly conserved. Of the
suspected acute encephalitis patients from September 2016-August 2018. CSF was
two amino acid mutations found at positions 29 and 57 in Indian isolates, the
collected from 233 patients, serum from 66 patients and both CSF & serum was
mutation F57V is part of a T-cell epitope.
obtained from 58 patients. These samples were subjected for detecting anti-JE
Keywords: HPV16, E7, Phylogeny and evolution, Bioinformatics, Structure-function
IgM antibody employing IgM antibody capture ELISA (MAC-ELISA) supplied by
relationship.
National Institute of Virology (ICMR), Pune. The epidemiological and clinical profile
of the patients were noted in detail.
Study of Chikungunya virus infection in the state of Meghalaya
Results: In a total of 357 suspected acute encephalitis patients 87 (24.4%) were
Sapam S1, Phukan AC2, Prasad AK3
confirmed to be suffering from JE. Among the confirmed cases, anti-JE IgM
antibody was found in 49 (56.3%) CSF samples and 21 (24.1%) serum samples. In
Department of Microbiology 1,2,3, North Eastern Indira Gandhi Regional Institute of
17 (19.5%) cases both CSF & serum samples were found to be positive. Male
Health and Medical Sciences, Shillong, India
preponderance (64.4%) was seen among the infected patients where majority
( 61%) of patients were below 15 years of age. Infection was mostly predominant
Email: [email protected]
(95.4%) during the rainy season (June-October). Among confirmed cases fever
(95.4%), altered sensorium (51.8%), seizure (32.9%),coma (22.9%),headache
Key words: CHIKUNGUNYA, MEGHALAYA, MAC-ELISA
(25.3%) and neck rigidity (6.9%) were the major clinical findings. Most positive
BACKGROUNDS: Chikungunya is becoming a major health problem in North-East
cases were from the state of Meghalaya (68.9%; mainly from East Khasi Hills & Ri-
India and its prevalence is gradually increasing in this part of the country. This study
Bhoi Districts) followed by the neighbouring states of Arunachal Pradesh
was undertaken to detect the prevalence of chikungunya in patients attending a
(17.2%) and Assam (10.3%).
tertiary care centre in North east India and also to study the demographic and clinical
profiles of such patients.
Conclusion: Information generated from this study may be used for further
MATERIALS AND METHODOLOGY:This was a 1year study conducted from January
analysis of molecular epidemiology with characterization of the circulating viral
2017 to December 2017. Whole blood was the sample collected from suspected cases
strains in various parts of the NE region. Further the study will be helpful for
(as per the selection criteria mentioned in the National guidelines for Clinical
designing suitable control and prevention strategy in this region.
Management of Chikungunya, 2016). Serum was separated from these samples and
MAC ELISA was employed to detect anti-Chikungunya IgM antibody. The
Development of single tube multiplex Reverse Transcriptase Loop Mediated
demographic and clinical profile of the patients was noted in details.
Isothermal Amplification Method for simultaneous detection of West Nile
RESULTS:Serum sample of 47 out of 307 suspected cases were confirmed to be
&Japanese encephalitis virus.
suffering from Chikungunya. A whooping 66 (21.5%) samples was found to be give
equivocal results. These patients with equivocal results were asked to give repeat
Email:[email protected]
samples after 10 days but none of them followed up. Male (57.45%) preponderance
was seen among the infected patients where 21-30 years age group (29.7%) was
Background:West Nile virus (WNV) is an arthropod-borne virus that belongs to the
found to be predominantly affected. Majority of the cases were from Meghalaya
genus Flavivirus of the family Flaviviridae and is a member of the Japanese
(93.6%) followed by the adjoining state of Assam (4.25%). The most affected district
encephalitis virus (JEV) serocomplex that includes JEV. West Nile virus (WNV) and
in Meghalaya was West Garo Hills (93.1%). Most of the cases presented within 1- 5
Japanese Encephalitis virus are neurotropic Flaviviruses that have emerged globally
days since the onset of symptoms (78.72%). Among the confirmed cases fever
as a significant cause of viral encephalitis. Currently, there are no effective therapies
(85.10%), myalgia (72.34%), polyarthralgia (31%), headache (46.80%) and skin rashes
or vaccines against WN and JEV infection for use in humans. Therefore, the
(4.25%) were the major clinical findings.
prevention of WNV and JEV invasion is an important public health concern in regions
CONCLUSION: Information generated from this study may be used for further study
that have close links with areas in which WN and JEV is endemic.The early
in depth the molecular characterisation of the circulating strains of the viruses in the
confirmatory diagnosis of WNV and JEV infections are important for timely clinical
NE India. Further the study will be useful to detect any new emerging endemic area
management and epidemiologic control in areas where multiple Flaviviruses are
of chikungunya and to adopt preventive strategies in districts where Chikungunya
endemic. Present study was aimed to develop (mLAMP) assay targeting env gene for
cases are reported.
detection of West Nile and Japanese Encephalitis virus. We therefore established a
multiplex real- time Loop Mediated Isothermal Amplification assay to simultaneous
Study of Japanese Encephalitis viral infection among the patients attending tertiary
detect and distinguish West Nile and Japanese Encephalitis RNA in single reaction.
care centre in North-east India
Two target sequences, one specific for West Nile and the other specific for Japanese
Encephalitis virus were amplified by specific LAMP primers in the same reaction tube.
Biswas D1, Phukan AC2, Prasad AK3
After amplification at 650C for 60 minutes the amplified products were subjected to
melting curve analysis and thus could be distinguished based on the different melting
Department of Microbiology1,2,3, North Eastern Indira Gandhi Regional Institute of
temperatures (Tm values) of the two specifically amplified products.
Health and Medical Sciences, Shillong, India
Biocontrol of cholera using Vibrio phage Vmj1 in fresh water microcosm of North
Email: [email protected]
india

Key words: JAPANESE ENCEPHALITIS, MAC-ELISA, NORTH-EAST INDIA

Naveen Chaudhary1, Balvinder Mohan2, Neelam Taneja3
Background: Acute encephalitis syndrome caused by Japanese Encephalitis (JE) is
a major public health problem of North East (NE) India. The present study was Department of Medical Microbiology1,2,3 PGIMER, Chandigarh-160012, India
undertaken with the objective to study the prevalence of Japanese Encephalitis
among the suspected patients attending tertiary health care centre in NE India and
NT conceptualized the study. NC and BM conducted experiments and obtained data. Results and conclusion:Out of the 132 samples collected 60 samples turned out to
NT, NC analysed data and wrote the manuscript. be positive for Enteroviruses by qRT PCR targeting the 5’UTR region. The conventional
PCRs showed positivity for EV71 -13, CV6 – 19, CV16-1, and co-infection of EV71 and
Email. Id:[email protected] CVA6 in 10 cases respectively. Sequence analysis revealed few variations and the
strains obtained from our study was closer to South East Asian strains.
Background:Biocontrol of cholera using Vibrio phage Vmj1 in fresh water microcosm
of North India. Cholera remains an important public health problem in India. Cholera Norovirus infection and disease in a cohort study in Vellore, India
outbreaks frequently occur in our region in North India One-third of India’s
population lives under the threat of cholera. Vibrio cholerae serogroups O1 is the Maheswari K, Hanusha J, Nirmal Kumar, Sidhartha Giri, Gagandeep Kang
currently associated serogroups with epidemic and pandemic cholera. Cholera
outbreaks frequently occur in our region in North India .In our previous work on Department & Institution: Wellcome Trust Research Laboratory, Christian Medical
ecology of V.cholerae in fresh water environments, we could demonstrate 11.1 % of College, Vellore
water samples from outbreak affected areas to be positive for V. cholerae O1.We
collected 117 environmental samples including water and sewage samples from Background:Norovirus are the second most cause of acute viral gastroenteritis after
cholera-affected areas during the time of outbreaks, viable but nonculturable rotavirus infection in all age groups worldwide. Majority of the studies on norovirus
(VBNC)V.cholerae O1 was demonstrated in 40.69% samples from cholera-affected associated diarrhea are hospital-based and very few are community-based studies.
areas .It has been previously demonstrated that cholera stools contain lytic Objective: To compare the prevalence of norovirus infection in symptomatic and
vibriophages and they also are important in environmental biocontrol of cholera. asymptomatic stool samples collected from children from birth up to the age of 3
Forty five samples of community sewage water and drinking water from five years in Vellore.
different cholera affected sites in North India including Manimajra, Ramdarbar, Methods: A total of 1711 diarrheal and 1400 control stool samples were tested from
Chandigarh Ludhiana and Ambala were collected Bacteriophage isolation and a birth cohort of 373 children, followed up for 3 years during 2002-2006. In the cohort
purification was done by filtration and ultracentrifugation. We isolated total five study, surveillance stool samples were collected once every two weeks during the 3
vibriophages (11.1 %) acting against V. cholerae O1 Ogawa serotype Out of five,4 years of follow up. For each diarrheal episode, we selected a non-diarrheal/control
phages were isolated from sewage (Ramdarbar ,Manimajra , Ambala,Ludhiana ,one sample from the same child with no diarrhea in the +/- 2 weeks period. All the
phage from each site). One phage was isolated from drinking water (Ludhiana). Only samples were screened for norovirus genogroups GI & GII using real time RT-PCR
one phage (VMJ1) could be propagated which was further characterised.by (qRT-PCR) targeting ORF 1 and 2 region of the genome. The samples positive for
transmission electron microscopy and SDS –PAGE. norovirus GI and/or GII by qRT-PCR were genotyped by conventional PCR followed by
We studied the killing efficiency of VMJ1against V. cholerae O1 Ogawa (isolated from Sanger sequencing.
a recent outbreak of 2015) in Sukhna Lake fresh water microcosm.Phage VMJ1 and Results:Of 1711 diarrheal episodes, 19.1% (327/1711) stool samples were positive for
V. cholerae O1 were grown in continuous culture by using lab scale fermenter. norovirus (56 GI, 263 GII, 8 mixed). 59.4% (38/64) of samples positive for GI, and
Growth kinetics was studied by using spectrophotometer and readings were taken 58.3% (158/271) of samples positive for GII were genotyped. Of the 1400 control
after every 5 minutes for 22 hrs automatically. A significant reduction in the Vibrio samples, 18.4% (258/1400) stool samples were positive for norovirus (51 GI, 197 GII,
growth was seen as compared to control (p<0.01) suggesting that this phage may be 10 mixed) by qRT-PCR. A total of 145 (54.1%) qRT-PCR positive control samples were
a good candidate for biocontrol of V. cholerae infection. genotyped. GI.3 and GII.4 were the most common genotypes in both diarrheal and
control samples.
Molecular Mapping of Enteroviruses causing HFMD from South India Conclusion: In this reports, this study re-emphasizes high rate of symptomatic as well
as asymptomatic norovirus infections in the community, and found GI.3 and GII.4 as
Ferdinamarie Sharmila P1, Christi R1, Rahul Dhodapkar1 the most common strains circulating in the community.
Role of Viral Hemorrhagic Fever (VHF) laboratories network in strengthening of VHF
Department of Microbiology, JIPMER Puducherry1 surveillance in India

Background:Hand, foot and mouth disease (HFMD) is a communicable disease Yadav PD, Sahay RR, Shete AM, Pradip Barde, Amita Jain, Sapkal GN, Bharti Malhotra,
affecting infants and children and rarely adults. It is an acute viral infection caused by Kakru DK, P Vijayachari, Bhagirathi Dwibedi and Mourya DT
a group of Enteroviruses, including Coxsackievirus A16 (CA16) and Enterovirus 71
(EV71). The disease is usually mild characterized by fever, painful sores in the mouth, ICMR-National Institute of Virology, Pune, Maharashtra, India
and a rash with blisters on hands, feet and also buttocks. Infection with EV71 can
sometimes cause mortality in children. Email- [email protected]
In recent years the incidence has increased in South East Asia including India. HFMD
was first reported in India from Calicut (Kerala) in 2003, thereafter outbreaks were Keywords: VHF, CCHF, KFD, Zika, sensitivity
reported from Maharashtra, Assam, West Bengal, Odisha, Rajasthan, Jammu and Background: Viral Hemorrhagic Fever (VHF) diseases caused by RNA viruses
Kashmir. Not much reports are available from Tamilnadu & Pondicherry, hence this belonging to the families; Filoviridae, Arenaviridae, Nairoviridae and Flaviviridae.
study. Emergence and re-emergence of VHFs are major public health concern. Under Global
Mass polio vaccination has eliminated polio viruses from the gut, thereby increasing Health Security Agenda (GHSA), ICMR-NIV, Pune is continuously working in
the chances of coxsackie viral and echoviral infections. This could be one of the collaboration of Centers for Disease Control and Prevention (CDC), Atlanta, USA since
possible reasons for the increased incidence especially in India. 2015. Multisite six VHF laboratories network were established in India to understand
Materials and methodsA total of 132 cases were identified and included from the the presence of Kyasnur Forest Disease virus (KFDV) and Crimean Congo Hemorrhagic
dermatology and Venereology clinics JIPMER. Throat swab specimens were used for Fever virus (CCHFV) in the region where the presence is not known and also to screen
RNA extraction. qRT PCR was done to confirm the presence of Enteroviruses following the other VHF causing viruses like DENV, CHIKV and ZIKV.
which conventional PCR was done for EV71, CV6, and CV16 separately targeting the Methods: 6479 Acute Febrile Illness (AFI) cases enrolled from September 2016 to
VP1 region. Sequencing was undertaken to identify the presence of any mutation and December 201. Tested for DENV, CHIKV and ZIKV. Negative samples were tested for
to build a phylogenetic tree using MEGA. CCHFV and KFDV. SPSS and Epi info software were used for analysis.
Results: Positivity observed for DENV (16.48%), CHIKV (8.52%) and dual positivity Priya Hemavathy. R, Archana Sriraman, Rajesh.A, Sidhartha Giri, Gagandeep Kang
(2.75%). No positivity for ZIKV. Negative samples were tested and found to be
negative for CCHFV and KFDV, but looking at small denominator of tested samples, it Department & Institution: Wellcome Trust Research Laboratory, Christian Medical
will be difficult to say that no presence was detected from these centres. Diagnosis College, Vellore
of DENV and CHIKV is mainly based on the clinical case definitions of suspected,
probable & confirmed cases given by NVBDCP. Screening of DENV & CHIKV positive Background: Diarrhoeal diseases are a major cause of hospitalizations and child
cases had increased to 789 with 63.32% sensitivity and 644 with 88.22% sensitivity deaths globally. Of India’s 78,000 rotavirus associated annual deaths of children,
respectively by new proposed case definition. Varying seasonal trends for CHIK and about 59,000 occur in the first 2 years of life.
DEN were observed. Aim and Objective: To compare the prevalence of rotavirus associated diarrhea in
Conclusions: Study provide VHF network based prevalence of DENV/CHIKV. KFDV children <5 years of age in the outpatient and inpatient settings in Christian Medical
and CCHFV screening on non-dengue, non-chikungunya cases helped in College (CMC), Vellore, over a four years period (July 2012- June 2016), and also to
strengthening the surveillance. The capacity building of these six biomedical evaluate the genotype diversity of rotavirus causing diarrhea in the two settings.
laboratories had helped in preparedness of country. Slight modification in standard Methods: Stool samples were collected from the enrolled children (under five age
case definition, higher sensitivity was achieved for screening of DENV and CHIKV. group) from inpatient and outpatient settings of CMC Vellore, after obtaining
informed consent from parents/guardian. The samples were initially screened for
Clinically significant substitutions for genotype specific Direct acting antivirals rotavirus VP6 antigen using commercial Enzyme Immunoassay (Rotaclone) kit. For
the EIA positive samples, genotyping was performed to determine the G (VP7) and P
Poonam Kanta, Varun Chauhan, Mini P Singh. (VP4) types using conventional RT-PCR.
Results: A total of 1337 children from the inpatient setting and 775 children from the
Department of Virology, Post Graduate Institute of Medical Education and Research, outpatient setting, who had provided a stool sample, were included in the four years
Chandigarh, 160012 data analysis. Of the 775 children from the outpatient setting, 91 (11.7%) were
positive for rotavirus by EIA, compared to 29.3% (392/1337) rotavirus positivity in the
Introduction Recent development of potent direct antiviral (DAAs) for HCV have inpatient setting. The common rotavirus genotypes in the outpatient setting were;
revolutionized the treatment of Hepatitis C with > 90% patients achieving Sustained G1P[8] (48.4%), G2P[4] and G9P[4] (13.2% each). In the inpatient setting, G1P[8]
Virological Response (SVR 12) with these newly emergent pangenomic antivirals. (62.6%), G2P[4] (8.4%), and G9P[4] (8.2%) were common. Mixed genotyped were
However, recently resistant associated substitutions (RASs) have been observed to found in 5.5% and 7.9% in the outpatient and inpatient settings respectively. Partially
occur which are significantly correlated with the failure of treatment. In such scenario typed and untyped samples comprised 5.5% and 3.3% in the outpatient setting,
it will be pertinent to explore the exact mechanism of interaction patterns between compared to only 1.1% and 1.8% in the inpatient setting respectively.
the available DAAs and various known mutants of NS5A protein to answer the Conclusion:Rotavirus associated diarrhea was 2.5 times more common in the
mechanism for their development of resistance to DAAs. The aim of present study inpatient settings compared to outpatient, suggesting that rotavirus associated
was to determine the difference in the interaction patterns of available DAAs and diarrhea causes more severe episodes resulting in hospitalization.
various combinations of RASs in NS5A protein well known for genotype 1 (1a and 1b).
GFP tagging enables expression of Human T cell lymphotropic virus-1 p30 protein in
Material & methods All the reference sequences were obtained from nucleotide bacterial system
database of Pubmed (https://www.ncbi.nlm.nih.gov/pubmed/nucleotide) for HCV
genotype 1 (1a and 1b).The well known hot spots which are clinically correlated with Namdev P1and Anupam R1*
the resistance for Ledipasvir in NS5A were selected (amino acid position 28, 30, 31,
58,93) for genotype 1 and the reference was changed to mutant type according to Department of Biotechnology1, Dr. Harisingh Gour University, Sagar (M.P.), India.
the genotype specific RASs such as M28A/T/V/G, Q30/K/E/L/H/R, L31I/M/V, H58D,
Y93F/H/N/C for genotype 1a, L28M, R30G/H/Q, L31F/I/M/V, P58A/L/S/D, Y93C/H/I/R Email: [email protected]
for genotype 1b. The models were generated for the wild type and mutants using
Galaxy WEB online web server. Further the models generated were refined and the Background:Human T lymphotropic virus type -1 (HTLV-1) is a complex
quality of the models were checked by Ramachandran Plot analysis.Interacting deltaretrovirus and the only human malignant retrovirus. HTLV-1 is the causative
patterns were studied with three DAAs (ledipasvir and velpatasvir) and wild and agent of an aggressive malignancy called Adult T Cell Leukemia (ATL). pX region of the
mutant proteins using molecular docking approach. viral genome encodes an accessory protein p30 which is reported to be involved in
promoting the viral spread, persistence and also in viral gene regulation at the
Results The Ramachandran plot analysis revealed that the models generated were of transcriptional and post transcriptional levels. p30 competes with Tax for p300
good quality showing almost more than 90% of the amino acids in favored region. binding at transcriptional level and involved in nuclear retention of tax/rex mRNA to
Further the molecular docking analysis revealed significant differences in the free curb the viral gene expression promoting latency. p30 modulate cellular environment
energy of the NS5A gene on acquiring such mutations, thus conferring resistance to to aid cellular transformation by binding various host proteins such as ATM, REGγ,
these drugs. Among the hotspots included in the present study, the amino acid PRMT5 and by altering DNA damage repair mechanisms, cell cycle regulation.
substitutions at position 58 and 93 were identified to be most significant in terms of However, the low expression level and instability of p30 in vivo have been major
energy change in the 3D model of the protein. hurdles in studying its structure-function relationship in the context of HTLV-1
pathobiology.
Conclusion The study provides an in depth understanding of drug resistance Materials and methods:HTLV-1 p30 protein expression in large quantities was carried
mutations in the NS5A gene of the HCV genotype 1 and will further help the out in E. coli using a bacterial expression vector pRSET/EmGFP which expressed p30
researchers in developing an effective strategy for HCV treatment. by itself (Swap clone) and p30-GFP fusion protein (Insert clone). The expression was
tested by immunoblotting. MALDI-TOF mass spectrometry analysis was performed to
confirm the identity of p30.
Comparison of rotavirus associated diarrhea in children <5 years of age in an Results:Expression studies of the p30 protein led us to conclusion that fusing GFP
Outpatient and Inpatient Setting in Christian Medical College, Vellore, India stabilizes p30 and enables its expression in bacterial system. However, the expressed
p30 is going into insoluble fraction. SDS and urea were used to solubilize p30 and was
purified for the first time using the His-tag affinity chromatography. The mass healthy controls than the patients with chronic Chikungunya (52.14% vs. 32.51%) with
spectrometry analysis confirmed the identity of HTLV-1 p30. a p-value 0.003, suggesting that subjects with showed AA genotype were protective
Conclusion:Large amounts of p30 protein will enable better insights into host protein from chronic Chikungunya indicating their resistance from progression to
interactions, molecular and cellular roles of p30 in HTLV-1 life cycle and pathogenesis Chikungunya chronicity. The frequency of GG genotype was more in patients with
which would be instrumental to develop novel treatments for ATL. chronic Chikungunya than the healthy controls (26.99% vs. 11.65%) with a p-value
0.0005 indicating the susceptibility of the genotype. Conclusion: Association of GG
Distribution of rotavirus genotypes in Indian children <5years of age hospitalized genotype of NKG2A gene as a susceptible gene and the emergence of AA genotype
for diarrhoea as a resistant gene towards Chikungunya virus infection is being reported for the first
time. Our results suggest that genetic susceptibility and/or resistance to Chikungunya
Email:[email protected] infection may be modulated by genes coding NK cell inhibitory receptors.

Divya P, Priya Hemavathy R, Archana Sriraman, Sidhartha Giri, Gagandeep Kang Hepatitis E virus seroprevalence among blood donors in Pune, India

Department & Institution: Wellcome Trust Research Laboratory, Christian Medical Tripathy AS1*, Puranik S2, Sharma M1, Chakraborty S1, Dekate U2 1
College, Vellore
Hepatitis Group, National Institute of Virology, Pune,Sus Road, Pashan, Pune,
Introduction: Rotavirus causes an estimated 11.37 million episodes of acute Maharashtra, India Department of Pathology, B.J. Medical College and Sassoon
gastroenteritis (AGE) in children <5 years annually in India, and approximately 78,000 General Hospitals2, Pune, India
deaths. The diversity of rotavirus genotypes causing diarrhoea varies across
geographical regions. E-mail: [email protected],[email protected]
Aim: To compare the genotype diversity of Group A rotavirus causing diarrhoea in
children < 5years of age in north and south India during a four year multisite hospital Background: Blood transfusion is a recently reported route of HEV transmission. It is
based surveillance study from 2012 to 2016. a bigger concern in regions where large scale HEV genotype 1 infections occur causing
Methods: Stool samples were collected from children <5 years hospitalized with more severe disease. The present study aims to assess the prevalence and rate of
diarrhoea from seven sites across India during 2012 to 2016, which included 5 HEV infection in the blood donors of Pune, India.
southern (Vellore, Trichy, Kolenchery, Hyderabad, Tirupati) and 2 northern (Delhi, Materials&Methods: A total of 2447 healthy blood donors were screened for anti-
Ludhiana) sites. Samples were collected after obtaining informed consent from HEV IgG and IgM antibodies. Anti-HEV IgM antibody positives were further subjected
parents/guardian. All stool samples were screened for group A rotavirus by ELISA to ALT measurement, HEV RNA detection, viral load quantification and phylogenetic
(Rotaclone). The EIA positive samples were genotyped by reverse-transcription analysis.
polymerase chain reaction. Results: Anti-HEV seroprevalence rate was 17.70%, while IgM prevalence rate was
Results: Of the 5834 samples from the 7 sites during the four years surveillance, 2069 0.20%. An age dependent increase in IgG seropositive rate was observed. Two of 5
(35.5%) were positive for rotavirus by EIA. Genotyping was performed for 2010 IgM-positives tested positive for HEV RNA. The viral load ranged from 3.5 x 104 - 4.6
(97.1%) samples. G1P[8](56.3%), G2P[4](9.1%), G9P[4](7.6%), G9P[8](4.2%), and x 105 copies/ml and belonged to HEV genotype 1.
G12P[6](3.7%) were the common genotypes in southern India and G1P[8](36%), Conclusion: HEV prevalence rate of 17.70% in the blood donors of Pune, India, a
G9P[4](11.4%), G2P[4](11.2%), G12P[6](8.4%), and G3P[8](5.9%) in northern India. developing country, goes at par with the developed countries. Current data of 0.20%
Mixed genotypes were more common in the north (10.6%) than the south (6.6%). The [5 of 2447] blood donors positive for anti-HEV IgM and 2 of them being HEV RNA
proportions of partially typed and untyped samples were more common in the positive suggest a need for consideration of cost-effective evaluation towards pooled
southern sites (1.3% and 2.5% in the south, compared to 0.5% and 1.2% in the north HEV RNA testing in blood banks. Key words: Hepatitis E virus, blood donors, age,
respectively). seroprevalence, western India
Conclusion: The study highlights the high burden of rotavirus gastroenteritis in India
and the diversity of rotavirus genotypes across different geographical regions. Role of antigen-antibody combination ELISA in the diagnosis of Hepatitis C virus
disease
Association of NKG2A Gene Polymorphism (A>G) in chronic Chikungunya infection
in Western Indian population Jayakar S1, Dhodapkar R2, Hamide A3, Parameshwaran S4,

Anuradha S Tripathy, Mohini A Ganu, Sonam Lata, Sneha Senapati, Department of Microbiology1,2 Department of Medicine3,Department of Nephrology4
JIPMER, Puducherry,India
Hepatitis Group, ICMR-NIV, Pune
Email:[email protected]
Email:[email protected]
Keywords:HCV, CKD,Acute hepatitis,Antigen-Antibody ELISA
Key words: NKG2A, Genetic Polymorphism, Chronic Chikungunya. Aims & Objectives:To assess the diagnostic yield of “Antigen-Antibody” combination
Background: Studies on humans have shown the participation of NK cells in the early ELISA assay versus the antibody only ELISA assay in detection of HCV among Acute
control of Chikungunya, association of higher NK cell inhibitory receptors expression Hepatitis patients and Hemodialysis patients.
and lack of NK cell functionality with chronic Chikungunya infection. Hence we aimed Background:Currently available antibody detection assays have a long window period
to investigate an association between NKG2A (a NK cell inhibitory receptor) gene and detects Hepatitis C virus infection only when IgG antibodies are present.Antibody
polymorphism (A>G) and chronic Chikungunya infection in western India. Methods: assays also have a high false reactivity.Hence the need of an improved diagnostic kit
one hundred and sixty three patients with chronic Chikungunya and 163 Chikungunya in early detection of the hepatitis C virus infection.
negative healthy controls from western India were studied for NKG2A gene Materials&Methods:Serum samples were collected from patients with Acute
polymorphism (A>G) by PCR and RFLP methods and analysis was done using SPSS Hepatitis and End Stage Renal Disease (ESRD) on Hemodialysis. The samples were
software. Results: The SNP rs2734440 in NKG2A was associated with an increased subjected to “Antibody”(3rd generation ELISA) and “Antigen-Antibody”
susceptibility risk for chronic chikungunya.The AA genotype was more frequent in combination(4th generation ELISA) assay using commercial kits.
Statistical analysis:Positivity of Hepatitis C by using combination “Antigen- present study was aimed to characterize the circulating MuV strains in Dibrugarh
Antibody”(4th gen ELISA) and only Antibody assay (3rd gen ELISA) is expressed as district of Assam, India during 2016 and 2017.
proportion with 95% confidence interval.Comparison of proportion of positivity Material & methods: A total of 66 (throat swab/blood) samples were collected from
between combination “Antigen-Antibody” assay and only Antibody assay was done patients who presented with fever and unilateral/bilateral parotitis. Blood samples
using Chi-square test among CKD and viral hepatitis patients. (n=23) were investigated for IgM antibody against Mumps and molecular detection
Results:From the 417 samples collected, 243 belonged to CKD and 174 were acute was carried out in throat swab samples (n=66) utilizing TaqMan assay. Genotyping
hepatitis patients.Out of the 243 CKD patients 12 were positive by Antigen-Antibody through sequencing was done targeting the small hydrophobic (SH) gene and its
assay and 8 were positive by Antibody assay, Acute hepatitis group, 6 were positive genetic variations and phylogenetic analysis was performed.
by “Antigen-Antibody” kit while only 1 was positive by Antibody assay. None of the Results: IgM antibody was detected in 57% (13/23) cases and MuV RNA was detected
samples which showed positive by Antibody assay were negative by “Antigen- in 55% (36/66) cases, of which 58% were male and 42% were female. Majority of the
Antibody” assay. Among the CKD patients comparison of both kits showed a cases were children. Sequence and phylogenetic analysis of the SH gene revealed
difference of 0.017% with 95%CI of -1.5-1.6 (p=0.92) while acute hepatitis patients simultaneous circulation of Genotype C (76%) and G (24%) in the corresponding
had a difference of 5% with 95%CI of 1.1-9.6%(p=0.011). years.
Conclusion:The study states the higher diagnostic yield of Ag-Ab assay among acute Conclusions: To our knowledge this is the first report of circulating MuV strains from
hepatitis patients. North-east India. The study provides important genetic baseline data for the
development of prevention and control measures of mumps. Further bioinformatics
Hepatitis C Virus coded proteins modulate cellular metastasis suppressor Nm23-H1 analysis which is in process would shed more information on the genetic diversity of
and promote cell migration and invasion MuV and its evolution.

Lohit Khera1,2, Catherine Paul1, Rajeev Kaul3* In silico mapping of CD81-E2 interaction across HCV genotypes 1 and 3

Department of Microbiology,University of Delhi South Campus1,3 New Bhattacharjee C1,, Nandy S2, Das P1 and Mukhopadhyay Aa*
Delhi,Department of Molecular Cell Biology,Weizman Institute of Science2, Israel.
Department of Life Sciences, Presidency University1 Kolkata, India; Department of
Background:Hepatitis C Virus (HCV) is the major etiological agent of Hepatocellular Biochemistry & Biophysics, University of Kalyani2, Kalyani, India.
carcinoma (HCC) which is the third most common cause for cancer related deaths.
HCV induced HCC is a multi-step process which involves alteration of several of host Key words: HCV, CD81, in silico docking
regulatory pathways. One of the key features of HCV associated hepatocellular Category: Virus host interactions
carcinoma is the metastasis of cancer cells to distant organs. Human Nm23-H1 is one Introduction:Hepatitis C Virus is a blood borne pathogen responsible for chronic
of the best studied metastasis suppressor protein which has been shown to be hepatitis in more than 71 million people worldwide. Wide variation across strains and
modulated in many human cancers. Our work shows that HCV envelope protein E1 genotypes are one of the major hurdles in therapeutic development and treatment.
protein expression as well as HCV infection induces pro-metastatic effect on cancer While genotype 1 remains the most extensively studied and abundant strain,
cells which is simultaneous to Nm23-H1 transcriptional down-regulation and Nm23- genotype 3 is second most prevalent and more virulent.
H1 protein degradation. Moreover, Nm23-H1 intracellular localization is significantly Methods:We have compared differences in the glycoprotein E2 across HCV
altered in cells expressing HCV E1 protein. Importantly, overexpression of Nm23-H1 genotypes at nucleotide, protein and structural levels using various tools of
can rescue the cancer cells from pro-metastatic effects of HCV E1 and HCV infection. bioinformatics.
We further show that HCV capsid protein ‘Core’ can co-localize and interact directly Results and Discussion: E2 sequences from 29 strains across genotypes 1a, 1b, 3a and
with Nm23-H1 within the cancer cells. This interaction results in modulation of anti- 3b revealed a preference for C-richness which was attributed to a distinct bias for C-
metastasis properties of Nm23-H1. HCV core promotes Nm23-H1 protein rich codons in genotype 1 and to a lesser extent in genotype 3. Amino acid level
sumoylation, degradation, as well as transcriptional downregulation. Our study now comparison revealed an N-terminal region conspicuously conserved with majority of
provides evidence for role of HCV-Core and E1 proteins as pro-metastatic proteins the changes at the C-terminal half of the proteins. In silico models of E2 glycoproteins
which can modulate functions of cellular metastasis suppressor Nm23-H1 in HCV and docking analysis with the energy minimised PDB-CD81 model revealed unique
mediated cancer metastasis. interacting residues in both E2 and CD81. E2 of genotype 3a and CD81 had the
strongest interaction. In conclusion this is the first comprehensive study comparing
Circulation of two Mumps virus (MuV) Genotypes in Assam, India during 2016 and E2 sequences across genotypes 1a, 1b, 3a and 3b revealing stark genotype-specific
2017 differences which requires more extensive investigation.

Sarmah K, Borkakoty B, Sarma K, Bora PK Molecular and Sero-prevalence of Different Viral Infections in patients attending a
tertiary care centre in Meghalaya.
Regional Medical Research Centre, N.E. Region, Dibrugarh, Assam, India. Pin- 786001
Marbaniang Merinda Shylla Amberly, Nongsteng Ibakorlin, Marbaniang Risbunlang,
Email:[email protected] Prasad AK, Phukan A.C

Background: Mumps is a vaccine preventable disease and therefore Mumps vaccine Department of Microbiology North eastern Indira Gandhi Regional Institute of Health
has been used as a component of the trivalent Mumps-measles-rubella vaccine. and Medical Sciences & Principal Investigator of State
Recent reports suggesting the re-emergence of Mumps infection worldwide in the VRDL,Meghalaya,Shillong,India.
vaccinated populations signifies the declining efficacy of the vaccine. Waning of
immunity attributed to the variations between the circulating and the vaccine strains Email: [email protected]
remains one of the major cause of Mumps outbreaks. Moreover, Mumps is not
recognized as a public health problem in India due to lack of comprehensive reports Key words: VRDL, VIRAL INFECTION, PREVALENCE, MEGHALAYA, NORTH-EAST INDIA
on its infection, documentation of clinical cases and published studies though it ARBOVIRUS, HEPATITIS, ROTAVIRUS,Influenza.
causes high degree of morbidity and severe complications in young adults. The
Elucidating amino acid differences among dengue sero and genotypes and their
implication in dengue virulence Department of Biotechnology, Indian Institute of Technolgy,Roorkee, India

Bibhudutta Mishra, Dr Raviprasad Aduri,BITS Pilani K K Birla Goa campus. Goa, India Introduction:Chikungunya, an Alphavirus (genus of Togaviridae family) is enveloped,
arthropod borne, positive-sense single-stranded RNA virus. Re-emergence of the
Email:[email protected] Chikungunya occurs time and again across the world but till date, no therapeutic is
available to combat the Chikungunya virus. The capsid protein of Chikungunya is
Key words: Dengue virus, pathogenicity present at the N-terminus of the structural polyprotein. It consists of two domains:
Background: Dengue virus (DENV) is one of the most important mosquito-borne virus the amino terminus and carboxyl terminus domain. The amino terminus domain
causing dengue fever (DF) and severe dengue (SD). It exists in four different interacts with the viral genomic RNA and other capsid molecules which lead to viral
serotypes, DENV-1, 2, 3, and 4. Again these serotypes are classified in to different genome RNA encapsidation and nucleocapsid assembly. The carboxyl terminus
genotypes based on the E-NS1 gene sequence. One of the factors attributed to the domain possesses auto- proteolytic activity and is also involved in virus budding. Since
severity of disease is the intrinsic genetic composition of the virus. In this study we capsid plays a prominent role in the viral life cycle so it can be a potential target for
focused on how the differences in the amino acid sequence of structural and non- antiviral drug development.
structural proteins may lead to varying degrees of pathogenesis. Materials and Methods:In this study the capsid protein of Chikungunya virus has
Material and methods: Complete genomes of Dengue from NCBI are classified in to been cloned, expressed and purified using various chromatography techniques.The
serotypes/genotypes using Vipr database. A consensus sequence for each of the three dimensional structure of Chikungunya capsid protein was used for screening
proteins is generated using Clustal Omega. Amino acid variation within a given and identification of inhibitors from a small molecule library. The validation of
protein among the genotypes of a serotype is generated using in-house python screened compounds was done by using AutoDock tools 1.5.6.
scripts. The observed amino acid variation is then mapped into the experimentally Results and Discussion:The identified compounds were tested for inhibitory activity
determined structure of the proteins, wherever available or onto the homology against capsid protease using FRET based assay and inhibition kinetics has been done.
model of the protein. The biochemical validated compounds have been further used to study the antiviral
Results: Both the structural and non-structural proteins show high amino acid efficacy against Chikungunya virus in cell based culture. Our biophysical, biochemical
sequence conservation within a given serotype, however there is a marked variation and antiviral characterization of potential inhibitors of Chikungunya capsid will pave
between the serotypes (for example, the linker region of NS3). Most of the amino the path for the drug development against the Chikungunya disease.
acid variation observed is mainly conservative in nature (i.e. polar to polar).
Nevertheless, we observed the following changes more frequently than the others: Detection of Dengue parameters and their correlation with platelet count among
Aromatic (Y or F) – L; (2) D/E – N/Q; (3) N/Q – H; (4) A/S/G – T; (5) Y/F – H. Mapping patients suspected with platelet count among patients suspected with dengue
these differences onto the structures revealed that most of these changes are in the infection
loop regions that may play an important role in protein-RNA interactions.
Conclusion: The observed amino acid variation in the dynamic loop regions of E and Neelofar Malik1 Singh.S.2, Farooq.U.3
NS3 protein suggest possible role of these changes
Department of Microbiology Teerthanker Mahaveer Medical College & Research
Importance of Rab1a in HCV endocytosis Centre, Moradabad

Bhattacharjee C1, Adhikary S1, Mukhoapdhyay A1 Email:[email protected]

Molecular Virology Laboratory1, Department of Life Sciences, Presidency University, Key words: Dengue, NS1, IgG, IgM
Kolkata, India Background: Dengue infection, an acute febrile arboviral disease has become a major
public health concern. Annually, it affects up millions of people worldwide. An early
Email: [email protected] and accurate diagnosis of dengue in acute phase of illness is important for identifying
an epidemic and for initiation of therapy. Detection of the secreted NS1 protein is a
Key words: HCVpp, Clathrin mediated endocytosis, endocytic sorting, Rab1a. new approach that aid in early diagnosis. Platelet count is the only non -dengue
Introduction:Hepatitis C virus (HCV) is an enveloped RNA virus, belonging to the parameter that can support the diagnosis of the dengue shock syndrome and dengue.
family Flaviviridiae. It is the leading cause for chronic liver disease and hepatocellular This study was done to detect dengue parameters and correlate them with the
carcinoma. Viral entry is via different hepato-cellular receptors, especially tetraspanin platelet count.
CD81. Interaction between CD81 and viral glycoprotein E1E2 is followed by clathrin Material and method– The study was conducted in virology section of microbiology
mediated endocytosis in a time dependent manner. In our present work using GFP department at Teerthankar Mahaveer Medical college and research center,
labelled HCV pseudoparticles (HCVpp), we are investigating the time line of HCV entry Moradabad (U.P.) Serum samples were collected from clinically suspected dengue
into hepatocytes. Receptor mediated endocytosis via clathrin coated vesicles has cases and tested for NS1 Antigen,IgM and IgG antibody by using
been previously shown to be regulated by the small GTPases, Rab1a, particularly at immunochromatographic method. The platelet count was also recorded in all
the early endocytic sorting step. In this study, we investigated the role of Rab1a in samples.
HCV endocytosis using a Rab1a knocked down cell line. Results- Out of 100 sample tested total 52(52%) specimen were positive for one or
Materials and methods:In a time-dependent study, GFP and luciferase tagged HCVpp more dengue parameters and 48(48%) specimen were negative. Among 39(75%)
trafficking was assayed in Rab 1a knocked down cell line and compared to the specimen were positve only for NS1 antigen and 13(25%) were positive for IgM only.
parental cell line, Huh7. Entry events were studied via confocal microscope and Conclusion- Control measures along with rapid diagnosis is the key to effective
western blot. management of dengue infection. Ahigh percentage of NS1 antigen in the study
Results and Conclusion:Our results show the importance of Rab1a in HCV trafficking. population indicated new infection warranting an immediate need of control of
mosquitos and preventive measures.
Studies of Antiviral Molecules targeting Chikungunya virus specific serine protease
KSHV, A Journey To The Oncogenesis
Fatma B, Saini R, Sharma R, Kesari P and Tomar S
Tathagata Choudhuri* Conclusions:Prevalence of HIV, HBsAg and HCV among ANC cases represents their
prevalence among normal population. Such studies are needed to know the
Departmet of Biotechnology, Visva Bharati, Santiniketan, West Bengal, India prevalence of such diseases in our society which can help us to form the guidelines
for patient management.
Email:[email protected]
Effect of dicotyledonous plant extracts on Hepatitis C virus entry events
Key words: KSHV, PEL, Everolimus, 1, 25(OH)2 D3, Theaflavin,
Background:primary effusion lymphoma (PEL) is a rare form of B cell lymphoma with Avik Bardhan1, Aparna Mukhopadhyay2 Chayan Bhattacharjee3
lymphomatous effusions in body cavities. HIV infected patients and patients with
immune compression are mainly subjected to be infected by KSHV induced PEL. Presidency University, Kolkata 1,2,3
Despite of different available anti-cancerous remedies targeting the cellular and viral
mechanisms didn’t give any promising feedback with poor survival. In our recent Email:[email protected]
study, we have investigated the efficacy of different component on KSHV induced
PEL.
Methods: The anti-cancerous property of Everolimus, 1, 25(OH)2 D3 and Theaflavin Background: Hepatitis C virus (HCV), a positive stranded RNA virus of the Flaviviridae
has been tested against KSHV induced PEL cell line in a dose depended manner. Cell family is the causative agent of Hepatitis which is a major health concern. The modus
death analysis was done by flow cytometry and protein expressions were studied by operandi of the virus leading to its pathogenesis is not fully understood and as such
real-time PCR and western blotting. no pan genomic potent vaccines have been discovered. Although emphasis has been
Results: Everolimus has shown to downregulate the KSHV latent antigen expression given on herbal medicines, most of them are polymerase inhibitors affecting the virus
with concurrent blocking of lytic reactivation for active virus replication and also at the replication stage in the host cell.In India, the major problem lays in the fact that
inhibited latent antigen mediated constitutively active STAT-3 and NF-κB signaling. blood transfusion events lead to the transmission of HCV in several cases calling for a
From the co culturing experiment with immature dendritic cells, we have found the remedial strategy to block the virus at the entry step.Some of the common plants in
activation of dendritic cells with increase in surface expression of CD86 and HLA-DR. Kolkata were selected for this study that has one of its family members conditions
1, 25(OH)2 D3 induces both death of HSHV infected PEL cells and KSHV replication in with established literature of its effectiveness in blocking HCV at the entry step. Also,
p38 and VDR dependent manner. With the dose dependent treatment of Theaflavin, literature survey showed the presence of certain organic active compounds that was
the functional status of viral proteins is emphasized and apoptosis of normal cells is responsible for the entry inhibition.
decreased.
Conclusions: The present study with the anti-viral as well as anti-cancerous effect of Previously in the lab, the E2 glycoprotein was modeled de novo since no
those component was a step forward analysis of the KSHV associated PEL and have crystallization structure was available in any database. The binding interactions of the
demonstrated a path of viral antigen disruption, viral reactivation and apoptotic and active compounds of the selected plants were checked in silico with the de novo
therapeutic role in the virus associated cancer. This work helps us better modeled E2 glycoprotein using molecular docking tools. Also, GFP and luciferase
understanding with the host and viral molecular interactions and promising target tagged HCV pseudoparticles (HCVpp) were prepared in the lab to perform the entry
area of further research. assays and study the effects of the selected plant extracts on the HCVpp.
Bioinformatics tools were employed to estimate the amino acid residues responsible
Seroprevalence of Hepatitis B, Hepatitis C and HIV infection in tertiary care teaching for the binding interactions and hence develop an idea about the binding site
hospital in (U.P.) thermodynamics of the E2 glycoprotein with the active compounds that lead to entry
inhibition
Sakshi Vishnoi1,Dr .Sudhir Singh2, Dr.Umar Farooq3,
Co-infection of Dengue with Chikungunya and Typhoid in a tertiary care center of
Department of Microbiology 1,2,3
Teerthanker Mahaveer Medical College & Research North India
Centre, Moradabad( U.P )
Galhotra S, Grover P, Jindal N, Dhuria N.
Email:[email protected]
Department of Microbiology GGS Medical College and Hospital Faridkot.
Keywords: ANC, HIV, Hepatitis B and C.
Background: HIV, Hepatitis B and Hepatitis C are serious global health problem. The Email: [email protected]
HIV/AIDS epidemics are one of the largest public health problem of 21st century.
Hepatitis B and Hepatitis C infection are associated with chronicity leading to cirrhosis Key words: Dengue, Chikungunya, Typhoid, Co-infection
and further progress to Hepatocellular carcinoma. The risk behaviors and routes of Background: Acute febrile illness is very common in patients especially between the
transmission of these infections are identical and hence it is observed that many month of June and September. This is a common clinical syndrome of dengue,
patients are co-infected with one or two of these infections (HIV, Hepatitis B and typhoid, Japanese encephalitis, Chikungunya, Leptospirosis, influenza A and Malaria.
Hepatitis C). Early diagnosis and appropriate treatment prevent the transmission as Nowadays, Co-infection with two or more infectious agents is becoming a major
well as progression of the disease, further increasing life expectancy. This study was public health problem.
aimed to assess the prevalence of HIV, Hepatitis B and Hepatitis C infection in ANC Material& Methods: A total of 1145 samples tested for dengue NS1 from May 2017
women attending ANC clinic of tertiary care teaching hospital in western U.P. to Nov 2017 were analyzed for the presence of other febrile illness i.e. typhoid and
Material and Method:The study was conducted in serology section of microbiology chikungunya fever. Out of these, 260 samples were positive for NS1 Ag and these 260
department at Teerthanker Mahaveer Hospital & Research center Moradabad (U.P.). samples were studied for typhoid and chikungunya co-infection by TYPHIDOT Rapid
The serum samples received from the ANC clinic were included in the study. IgM Immunochromatographic test and IgM Capture ELISA respectively.
Results:A total 250 samples were tested from ANC cases for HBsAg, HCV and HIV. Out Results: Dengue NS1 Ag positivity was 22.7% (260/1145). Of these positive (260)
of total samples, 42(16.8%) were reactive. In which 31(12.4%) were reactive for HCV, samples, 11(4.2%) were positive with Salmonella typhi by rapid IgM test and 9 (3.4%)
9(3.6%) were reactive for HBsAg and 2(0.8%) samples were reactive for HIV. were positive for IgM antibody for Chikungunya. There was no patient which was
positive for all the three infections.
Conclusion: In our Malwa Region of Punjab dengue and typhoid co-infection exists, HSV-2, CMV, Haemophilus ducreyi and Treponema pallidum. Serum samples were
early diagnosis of typhoid infections in patient suffering from dengue fever is required collected for HSV and syphilis serology.
as early antibiotic therapy in the former leads to a favorable outcome, while dengue Results:Scrapings from the perigenital ulcer were positive by PCR for HSV-2, CMV and
as such has no specific treatment and is treated symptomatically. Repeated T. pallidum, but tested negative for H. ducreyi. Culture and serology for HSV 2 was
outbreaks of dengue, chikungunya/dengue co-infections are occurring in our region. positive. Both VDRL and TPHA for syphilis were positive.
So, in clinically suspected cases of dengue or chikungunya fever, it is advisable to test Conclusion:This case underscores the importance of maintaining a broad differential
for both viruses in this area. diagnosis for genital ulcers in immunocompromised patients. Any genital or
perigenital ulcer should be investigated to rule out CMV in an immunocompromised
Epidemiology of viral respiratory infection in Odisha, India patient as its recognition could reflect systemic involvement and significantly affect
patient care. Key words: Herpes Simplex Virus, Cytomegalovirus, Human
Jyotsnamayee Sabat1, Subhra Subhadra1, Bhagirathi Dwibedi2, Subrata K Palo1 Immunodeficiency Virus, Haemophilus ducreyi, Treponema pallidum.
Sanghamitra Pati1
Identification of mutations in Haemagglutinin gene of Influenza A H1N1 pdm09
VRDL, ICMR- Regional Medical Research Centre, Bhubaneswar, Odisha1,2,3,4,5 strain from 2016-2017 outbreak in India

Background:Acute respiratory tract infections (ARI) are an important cause of Akshatha R1, Dhodapkar R2,Ferdinamarie Sharmila P3 Sistla S4 Wyawahare M5
morbidity and mortality worldwide. Global burden of ARI has been estimated to be
94037000 DALYs and 3.9 million deaths (WHO 2002). The aetiology of viral ARI can be Department of Microbiology1,2,3,4JIPMER, Department of Medicine5, JIPMER, India
determined in 80%–95% of cases. This study reports the viral etiology of respiratory
illness in eastern India during 2010 to 2017. Email:[email protected]
Materials and Methods:Throat/nasal swabs from 3302 hospitalised symptomatic
individuals were collected from 40 different hospitals of Odisha. Nucleic acid was Keywords:Influenza A H1N1, mutations, HA, severity
extracted by column separation method and subjected to RealTime PCR for Background:Circulating influenza viruses are estimated by the WHO (WHO) to infect
qualitative detection of suspected viruses. 5–10% of the population every year. Studies have shown that epidemics are most
Result:It was observed that 20.8% cases were positive for pandemic H1N1 2009. commonly caused by Influenza A (H1N1) pdm09 (66%), B/Victoria lineage (24%) and
Other viruses detected were Flu A in 12% cases followed by RSV (5.7%), Adeno (4.2%), A (H3N2) (10%). Over the years certain mutations in amino acids at position 222 in
Rhino (3.9%) and Parainfluenza (3.9%). Among all cases, 25% were children below 5 HA gene has been associated with higher virulence and severe disease by increasing
yrs of age and viral etiology was found in 34% cases. In these children, RSV was the affinity to respiratory tract mucosa. This study, the first of its kind, was done to
detected in 9.6% followed by Adeno in 5% of cases. Among adults, the common find whether there were any similar or different mutations in the outbreak of 2016-
viruses detected were Pandemic H1N1 2009 in 16.43% followed by Flu A in 15.3%. 17.
Conclusion:RSV was the most common virus detected among children. Viruses like Materials &Methods:The current work was carried on archived Influenza A H1N1
Pandemic H1N1 2009, Flu A, RSV, Adeno and PIV has caused high morbidity in the positive samples (qRT method) from Regional Influenza Laboratory, JIPMER. RNA was
state. Also Corona, Human Boca Virus, HMPV etc were also detected for the first time. extracted using commercially available extraction kit (Roche High pure viral nucleus
Hence, a routine surveillance is essential to monitor the seasonality and true acid kit) according to manufacturer’s instruction. Conventional one step RT-PCR was
prevalence of these viruses to support vaccination advocacy in future. done using AgPath –ID kit and pre-published primers sequences targeting the HA
region. Positive PCR products were purified by ExoSAP-IT (GE Healthcare), and then
Herpes Simplex Virus Type 2 and Cytomegalovirus Perigenital Ulcer in an HIV subjected to Sanger sequencing using M13 primer with Big Dye Terminator Reaction
Infected Woman Mix. Sequencing results were analysed using the MEGA 7.0 Software.
Results&Conclusion:Hundred samples were retrieved and were classified into 63 ILI
Rawre J1Namdeo D2Rosaleen Das3, Khanna.N 4, Dar,L5,Dhawan,B 5. and 37 SARI cases respectively. Only 48 samples showed positive by conventional PCR
with a product size 890-976 KB, a possible reason being that most of the samples had
Department of Microbiology, AIIMS,New Delhi,Professor, Department of a high Ct value (>33) and/or possibility of degradation in RNA. By Sanger sequencing
Dermatology and Venereology4, AIIMS,New Delhi, Professor,Department of few variations were identified and phylogenetic analysis revealed strains close to
Microbiology5, AIIMS,New Delhi; Department of Microbiology5, AIIMS New Delhi South Asian population.

Email:[email protected] Antiviral drug identification by targeting nsP3-G3BP complex of Alphavirus

Background Genital herpes (GH) is the most common cause of genital ulcer disease Mahajan S1, Kumar R1, McInerney G M2, Tomar S1;*
(GUD) worldwide; its association with human immunodeficiency virus (HIV) is well
established. Cytomegalovirus (CMV) is an important opportunistic agent in Molecular Virology Laboratory, Department of Biotechnology, Indian Institute of
immunosuppressed states particularly in HIV infected, manifesting as ocular and Technology Roorkee, Roorkee, Uttarakhand,1 India.,Department of Microbiology,
visceral involvement. Though cutaneous manifestations are rare, there are growing Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden2
number of reports of concurrent Herpes Simplex Virus (HSV) and CMV infections in
genital and perigenital ulcers of immunocompromised individuals. Here, we report a E-mail: [email protected]
case of mucocutaneous HSV-2 and CMV infection in a 39-year old woman with
acquired immunodeficiency syndrome, who presented with a perigenital ulcer. Background: Alphaviruses are enveloped, 70 nm diameter, spherical and 40 nm
Materials and Methods Scrapings from the perigenital ulcer were collected in Viral isometric nucleocapsid particles. Alphavirus infection in humans is transmitted
Transport Medium (VTM) for HSV culture and polymerase chain reaction (PCR) for through mosquitoes. Major symptoms of infection in humans include fever, rash,
arthritis, encephalitis. The diverse interactions of viruses with the host cell and
interaction of viral proteins with host proteins effectively manipulate host cell RNA dependent RNA polymerase as an antiviral drug target for emerging
antiviral response and cellular processes. The host cells have also evolved antiviral alphaviruses
defense mechanisms which target viral RNAs and proteins. One such mechanism of
defense is stress granule formation which is a dynamic assembly of proteins Pareek A, Mudgal R and Tomar S
regulating mRNA translation during time of stress and reduces bulk protein synthesis.
Mechanism of stress granule (SGs) formation is not yet completely established but Department of Biotechnology, Indian Institute of Technolgy,Roorkee India
largely dependent on TIA1/R and G3BP(Ras-GAP SH3 domain-binding ) protein. Role
of G3BP in cells exposed to environmental stress and viral infections is well Background:Alphaviruses are arthropod borne viruses which belongs to Togaviridae
elucidated. Non-structural proteins of alphaviruses act as host defence modulator family. These viruses have emerged as a serious threat to human lives from past few
that functions by disrupting stress granules formed due to cellular stress response to years. Chikungunya virus (CHIKV), Sindbis virus (SINV), Venezuelan Equine
viral infections. This disruption mechanism is explained through nsP3-mediated Encephalitis virus (VEEV) etc. are well known members of this family. There is no
recruitment of human G3BP1 protein via two tandem FGDF motifs. Thus, FGDF-motif medication and vaccine to cure the disease caused by alphaviruses. Therefore,
of nsP3 is directly affecting SG formation and going around the host cell defenses. So development of antiviral agents specifically targeting the key viral enzymes is
we hypothesize that by targeting G3BP and designing pharmacophores that block necessary to control the viral diseases. Alphaviruses contain a genome of ~11.5kb size
G3BP interaction with the viral partner will lead to novel antivirals. Molecular docking in which the two-third part translates into the non-structural polyprotein and the
has been carried out using CDOCKER and AutoDock to predict the binding- other one-third part translates into the structural polyprotein. The non-structural
conformation of screened peptidemimetics to the G3BP. Human G3BP has been polyprotein is processed by viral protease to form four mature replication proteins
expressed using bacterial expression system, and purified using Ni-NTA column that form the replication complex in infected cells.The mature non-structural proteins
affinity chromatography. His-Tag has been successfully removed using Tev protease formed are nsP1-4 in which nsP4 functions as a RNA dependent RNA polymerase
followed by reverse Ni-NTA affinity chromatography. Purified protein has been (RdRp). nsP4, the viral RdRp uses positive-sense RNA genome and replicates its. In
crystallised and co-crystallisation with inhibitors is in progress. Antiviral studies of this study, RdRp genes of two different alpahviruses and number of their N-terminal
identified G3BP inhibitors show promising results. truncation constructs have been made for expression in bacterial expression system.
Sindbis virus (SINV) RdRp has been successfully purified using Ni-NTA affinity
Expression levels of toll like receptors in the culture supernatant of microglial and chromatography and size exclusion chromatography using AKTA purifier (GE
neuroblastoma healthcare). Preliminary studies indicate that alphavirus nsP4 in presence of nucleic
acid forms higher oligomeric states. Additionally, biophysical characterization of
Shukla M1Chaturvedi .M2,Dhole.T.N,3 purified protein by CD shows metal binding and nucleotide binding. Crystallization of
protein for atomic structure determination is in progress.
Department of Microbiology 1,3 Sanjay Gandhi Post Graduate Institute of Medical
Sciences, Lucknow U.P,Amity Institute of Biotechnology2,Amity Institute of In silico analysis to understand more prolonged viremia in Dengue Virus Serotype-
Biotechnology, India. 2

Email: [email protected], [email protected] All India Institute of Medical Science, Bhopal

Keywords: Japanese encephalitis, Email:[email protected]

Background: Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis
in Asia. There is no effective method to cure this deadly disease. Activation of various Background: Delayed viral clearance (viremia beyond 5 days) was found significantly
pattern recognition receptors provides a first line of defense by inducing IFN-α/β, higher among Dengue virus-2 (DENV-2) infected cases during molecular
proinflammatory cytokines and chemokines. These early responses are protective by characterization of DENV serotypes that circulated in Madhya Pradesh region during
activating and amplifying antiviral mediators, as well as recruiting leukocytes the 2016 outbreak (Unpublished data). In view of this, we performed an in silico
expressing antiviral function. However, the same mediators promote tissue damage analysis to investigate antigenic variability in DENV serotypes which might be
if not dampened in a timely manner contributing to higher ability for immune evasion and prolonged viremia.
Materials and Methods: We aimed to study the mRNA gene expression of TLRs 2, 4 & Methods:Genome-wide association was carried out with existing DENV sequences
9 in microglial cell line BV-2 & neuroblastoma cell line Neuro-2A by Quantitative Real from ViPR database. Immune evasion potential of infecting serotypes was
time PCR (qRT-PCR). qRT-PCR was performed for total cellular RNA extracted to ascertained by computing antigenic variability in B cell and Cytotoxic T cell (CTL)
measure mRNA expression for TLRs 2, 4 & 9 in the cell supernatant of BV-2 and Neuro- epitopes of all DENV proteins, using BepiPred software and IEDB analysis Resource,
2A infected with JEV. Infection and treatment schedule of BV-2 & Neuro-2A cells: The respectively.
BV-2 & Neuro-2A cells were plated in 6-well plates and cultured until 70-80% Results: We first computed the variability-coefficient for each position in DENV
confluence monolayers were obtained. After 18 to 24 hrs incubation in 10% DMEM, polyprotein of all four serotypes. We observed that the average variability in DENV-2
the cells were adsorb with JEV (108 pfu /mL) at an MOI of 5 for 1.5 hrs. After polyprotein was higher as compared to other serotypes (p<0.05). We also observed
adsorption, the unbound viruses were removed by washing with sterile 1X PBS. The that NS2a protein of DENV-2 displayed highest average variability among all the
cells were then incubated and mRNA for TLRs 2, 4 & 9 was carried out at 0, 3 , 6 , 12 proteins. Next, we computed the frequency of variability in the sequences of B cell
& 24 hrs respectively. Mock treated control cells received only 1xPBS Statistical epitopes and HLA-I binders which might be responsible for the evasion of humoral-
analysis was done using graph pad prism version 5. p value ≤ 0.05 was considered immunity and cytotoxic T-cell responses respectively. The number of variable B-cell
significant. epitopes were significantly different between serotypes and highest in DENV-2
Results: We are analyzing the data. The results will be discussed at the time of (p<0.001) for variability scores >4. Similarly, the total number of variable HLA-I
presentation. binders were significantly different (p<0.05) between the four serotypes and
Conclusions: This study will help to identify new targets for the therapeutic treatment maximum in case of DENV-2 at variability thresholds >4.
of JEV infection and may lead to the development of potential pharmacological Conclusion:This study reports the occurrence of maximum variability in B cell and T
targets. cell epitopes of DENV-2 among all serotypes, which is consistent with the more
prolonged viremia observed in infection with this serotype.
Molecular and Phylogenetic analysis of Chikungunya virus outbreaks in Central of all typed samples. The circulating pattern of SaV genotypes varied during the study
India during 2016, 2017 period with GII.1 being predominant in 2013 and 2014 and GI.1 in 2015-2018.
Conclusion:Sapovirus infection was detected in a considerable proportion of acute
Regional Virology Laboratory, All India Institute of Medical Sciences Bhopal, Saket gastroenteritis cases in the setting described with an increasing trend over the years.
Nagar, Bhopal-462020, India A wide diversity of sapovirus genotypes were identified in these cases.

Email:[email protected] Structure based In-silico identification of potential inhibitors of the prefusion form
of Nipah virus (NiV) fusion (F) glycoprotein
Background: Chikungunya is now endemic in most parts of India. Central India
witnessed Chikungunya virus (CHIKV) outbreaks in 2016 and 2017 with huge number Murali R., Swathi Sree V., Ganesh V., Bondili J.S., and Bhadra Murthy V.
of patients visiting our tertiary care hospital. The present report is a hospital based Genomics and Proteomics Group, Department of Biotechnology, Koneru Lakshmaiah
cross-sectional study in which serological and molecular investigations of patients Education Foundation (Deemed to be University), Vaddeswaram, Guntur, Andhra
suspected with CHIKV infection were undertaken. Further, mutational and Pradesh, India 522502 Email:[email protected]
phylogenetic analysis was conducted to study the genetic relatedness of the Bhopal
strains with other Indian strains and worldwide strains. Background:Nipah virus (NiV) is an emerging and deadly zoonotic pathogen in the
Methods: During 2016 and 2017 outbreaks, samples from Chikungunya suspected genus Henipavirus; Family: Paramyxoviridae. NiV cause acute and severe respiratory
patients were collected and tested for IgM and Anti-NS1 antibody by ELISA and viral illness in humans with a high case-fatality rate more than 75%. Pteropid spp. bats
RNA was detected by RT-PCR. Some of the RT-PCR positive samples were sequenced (flying foxes) are the reservoir hosts of NiV. Recent deadly NiV outbreak in Kerala,
for partial E1 gene and analyzed to identify the newly emerging mutations and India during June, 2018 claimed 17 lives has put India on a high risk zone for future
understand the phylogenetic relationship among 2016 and 2017 strains as well as NiV infections. No vaccines or therapeutic remedies are available to prevent or treat
with other Indian and worldwide strains. patients exposed to NiV. NiV particles are enveloped and its genome is un-segmented
Results: Phylogenetic analysis revealed the present strains to be of ECSA genotype. negative sense single-stranded RNA of ~18.2 kbp coding six proteins. The viral
Emergence of a variant strain was observed in the year 2016, which became the envelope consists of two transmembrane glycoproteins, fusion (F) and attachment
predominant strain in this region in 2017. The strains showed high similarity with (G) glycoproteins responsible for the virus entry into host cells. NiV G binds to ephrin
recent New Delhi strains of 2015 and 2016. The epidemic mutation A226V emerged B2/B3 receptor and triggers NiV F leading to viral entry. NiV F and G have been the
in 2006 outbreak was found to be absent in the current strains. Among the important major target of antiviral strategies to prevent viral entry.
mutations viz. K211E, M289V, D284E, I317V & V322A observed in the recent strains, In our study, computational and cell line studies are being carried out to identify
I317V is found to emerge very recently as it is present only in Bhopal (2016, 2017) potential NiV inhibitors. Structure based In-silico analysis involving 200,000
and New Delhi strains (2015, 2016). molecules of natural origin were analyzed against pre-fusion NiV F (PDB ID: 5EVM).
Conclusions: This study has identified unique mutation I317V in the E1 gene, which is Two regions on NiV F (a) aa 130-190 and (b) 450-490 critical for NiV prefusion to
present only in New Delhi strains till date. This study warrants continuous surveillance postfusion transformation was targeted. Schrodinger; other suitable bioinformatics
for mutations having epidemic potential thus aiding in prediction of future outbreak. software were employed. Based on ADME parameters; tautomer conformations;
PAINS; XP Docking and MDS (20ns), seven unique molecules have been identified with
Epidemiology and genetic diversity of Sapoviruses in cases of acute gastroenteritis high binding capabilities with at least three amino acids viz. N155; K167 and V158 of
in children less than five years, 2012-2018 NiV F. Co-transfection experiments with NiV F and G plasmids on HEK-293T, Vero and
CHO cells are being carried out to determine the extent of the inhibitory activity of
Blossom Benny1, Soumya Roobini1, B.Manohar2, Samarasimha Reddy1, Sidhartha Giri1 these molecules in arresting syncytia formation. Promising inhibitor molecules could
Gagandeep Kang1, Benjamin Lopman3, Ira Praharaj1 be used in antiviral therapy against NiV.
Wellcome Trust Research Laboratory, Division of Gastrointestinal Sciences1, Christian
Medical College, Vellore, Tamil Nadu,Sri Venkateswara Medical College,
Thirupathi2,Rollins School of Public Health, Emory University3, USA. Anti-HIV drug Efavirenz induces cytotoxicity by initiating mitochondrial membrane
depolarization
Keywords: Acute gastroenteritis, Sapovirus, Genetic diversity
Background:Diarrheal diseases are a major cause of childhood mortality and Mandal A1, Ganta K K1, Chaubey B1
morbidity in Indian settings, and viruses have been found to be major etiologic agents
in children < 5 years. In Indian settings, the etiologic role of sapoviruses in acute Functional Genomics Lab., Centre for Advanced Study1, Department of Botany,
gastroenteritis and their molecular epidemiology is not well described. University of Calcutta, 35, Ballygunge Circular Road, Kolkata, India
Aim:To study the trends in sapovirus prevalence and their genetic diversity in cases
of acute gastroenteritis among hospitalized children ≤ 5 years in a tertiary care facility Email:[email protected]
in south India over a period 2012-2018.
Methods:Stool samples from children ≤ 5 years hospitalized for acute gastroenteritis Background: Efavirenz (EFV) is a non-nucleoside reverse transcriptase inhibitor and
at a tertiary care centre in Thirupathi, south India, collected as part of a surveillance an active constituent of the highly active antiretroviral therapy regime. It has
for rotavirus disease from 2012 till February 2018 were included in the testing for significantly contributed in control and management of human immunodeficiency
sapovirus. Following viral RNA extraction, qPCR was performed targeting the ORF1 virus propagation. However, EFV administration has led to severe adverse effects,
region. All samples with positive amplification with the qPCR assay, with Ct ≤ 35 several reports highlighted the role of EFV in mitochondrial dysfunction and toxicity
underwent nested RT PCR targeting the capsid region followed by Sanger sequencing but the molecular mechanism has been poorly understood.
for genotyping of sapoviruses. Contigs were generated using Sequencher software Methods:In the present study, human hepatoma cells Huh-7.5 were treated with
and genotypes identified using NCBI BLAST. non-cytotoxic concentrations of EFV and parameters like cytotoxicity, mitochondrial
Results:Out of 1266 samples, 28.44% (360/1266) were positive for Sapovirus by transmembrane potential, mitochondrial morphology, cytochrome c release,
qPCR. We successfully genotyped 81% (183/227) with a Ct cut off ≤ 35. We detected mitochondria-mediated apoptosis, mtDNA, mtRNA levels and EFV distribution into
sapoviruses belonging to all genogroups GI, GII, GIV and GV known to infect humans. the mitochondrial compartment were evaluated to understand sequence of events
12 different sapovirus genotypes were detected with genotype SaV GI.1 found in 44% leading to cell-death in EFV-treated cells.
Results:EFV at its clinically relevant concentrations was significantly toxic after 48 and (H1N1) pdm 09 virus across all nodal centres in India. The purpose of this study is to
72h of treatments. The EFV-mediated toxicity is initiated with the change in Δψm evaluate the performance of HiScript One-Step RT-PCR master mix manufactured by
which triggers a series of events like the cytochrome c release, alteration in HiMedia Laboratories in comparison with AgPath-ID One-Step RT-PCR master mix for
mitochondrial morphology, mitochondria-mediated apoptosis and finally leading to diagnosis of influenza A (H1N1) pdm 09 virus.
cell death. We have further observed a decline in the total mitochondrial content Method: A total of 50 samples were studied (30 positive and 10 negative
after 48h of EFV treatment at IC50 concentration which was also reflected in reduced retrospective cases of influenza A (H1N1) pdm 09 virus along with 10 control samples
mitochondrial DNA and RNA levels. collected from healthy volunteer). Viral RNA was extracted using QIAamp Viral RNA
Conclusion:EFV being a lipophilic molecule internalized into the mitochondrial Mini kit. One-Step Real-time PCR was performed on RNA extracted from all
compartment which causes depolarization of Δψm and subsequently leads to a specimens in triplicates using both company master mixes. Applied Biosystems’s CDC
cascade of events to cell death. This study underscore mitochondrial dysfunction licensed Pandemic H1N1/09 Assay Set v 2.0 was used in this assay.
upon EFV treatment. Results: HiScript One-Step RT-PCR master mix on evaluation showed better sensitivity
with earlier detection of more than 1 cycle threshold (Ct) values as compared to the
Sero-prevalence of dengue fever in patients with acute febrile illness in a tertiary AgPath-ID One-Step RT-PCR master mix. In addition, the turnaround time for
care hospital diagnosis was 20 mins less using HiScript One-Step RT-PCR master mix in comparison
to AgPath-ID One-Step RTPCR master mix.
Dr Veenu Gupta Conclusion: The CDC protocol of real-time RTPCR for influenza A (H1N1) relies on
limited approval of Real-time master mixes for influenza A (H1N1) pdm 09 virus
Dayanand Medical College and Hospital Ludhiana diagnosis. Also, the availability and cost of these reagents is a critical element
especially in resource limited countries like India. The present preliminary study
Email: [email protected] provides laboratories with a more sensitive One-Step Real-time PCR master mix that
can be used as a master mix of choice during the routine testing and outbreak of
Background and objectives: Fever is the commonest presentation of patients seeking influenza.
healthcare in developing countries. The differential diagnosis for acute febrile illness
includes malaria, dengue, enteric fever, leptospirosis, rickettsiosis, and other Differentiation between primary and secondary dengue virus infection by IgM: IgG
infections. Dengue is one of the most common mosquito-borne viral infections. ratio
Dengue has emerged as a major public health concern in terms of mortality and
morbidity. The clinical and epidemiological profile of dengue infection changes from Charu1, Agarwal RK2, Mittal G3, Ahmad S4
time to time. Since there is no immunoprophylaxis or specific antiviral therapy
available, timely and rapid diagnosis plays a vital role in patient management and Department of Microbiology 1,2,3,Department of Medicine4, Himalayan Institute of
implementation of control measures. The study was conducted t to know Medical Sciences, Dehradun, India
seroprevalence of dengue fever among patients with acute febrile illness and to study
their clinical and laboratory profile. Correspondence: [email protected]
Methods: A total of 6705 patients with history of acute febrile illness admitted in the
hospital over a period of one year were enrolled. Serological tests like Dengue IgM Keywords: Dengue, Primary, Secondary, IgM, IgG, ratio
and NS1 antigen ELISA were performed to confirm the diagnosis. Serologically Background: Dengue exhibits varied clinical presentations and often unpredictable
confirmed patients of dengue fever were studied for their clinical presentation and clinical evolution & outcome. More than 70% (about 1.8 billion) of the world
lab parameters. population is at risk for Dengue. Secondary infection with the virus is an important
Results: The Seroprevalence of dengue fever was 17.3%. The most affected age group risk factor for the development of severe forms of the disease. Methods which
was 26-35 years with male predominance. Most dengue cases were seen in the discriminate primary and secondary DENV infection have prognostic importance. This
months of August to December. Common clinical symptoms and signs were myalgia study was undertaken to determine the best cut off point of IgM: IgG ratio in acute
(72.4%), arthralgia (34.4%), vomiting (50%), abdominal pain (38.6%), and phase sample and to determine the prevalent serotype of DENV in the study area.
hepatomegaly (27.4%). Lab parameters revealed leukopenia and thrombocytopenia Materials and Methods: A tertiary hospital based cross sectional study was carried
in most cases. Common complications were shock and encephalopathy. Hepatic and out and the records of 936 OPD and IPD patients suspected of dengue between Jan
renal failure was seen in 3.7%and 1.8% of patients respectively 2017 and Dec 2017 were collected. Dengue Duo rapid test and IgM Capture ELISA test
Conclusion: Tropical infections should be considered as important cause of acute were carried out on the blood samples. Of these, 234 tested positive for dengue
febrile illness. Dengue fever has a very non-specific and variable presentation. Due to either by NS1 antigen or IgM antibody testing. 91 random samples were stored at -
the overlapping clinical presentations, diagnosis must be confirmed by specific 20˚C and further processed for IgG antibody testing by ELISA, RT PCR and Nested PCR
diagnostic tests Keywords: Dengue fever, seroprevalence, clinical manifestations, was also performed.
acute febrile illness Results: Of the 9l confirmed cases, 51.6 % (47) turned out to be primary and 48.4 %
(44) secondary dengue infection. Mean IgG index and IgM: IgG index values were
Evaluation of HiScript One-Step RT-PCR master mix with AgPath-ID One-Step RT- significantly allied with dengue illness. Best cut-off of IgM: IgG ratio was found to be
PCR master mix for diagnosis of influenza A (H1N1) pdm 09 virus 1.59 with sensitivity of 85.11 %, specificity of 100 %, accuracy level 92.3 % and
negative likelihood ratio 0.15 for differentiating primary and secondary dengue. The
Gohil D1, Kothari S2, Subramanian A1, Todkar P2, Khadke K1, Naik N2, Warke R1 results of RT PCR indicated predominance of DENV 2 infection.
Conclusion: 1.59 as a cut off for IgM: IgG ratio is recommended, a ratio of ≥ 1.59 will
Department of Molecular Biology1, HiMedia Laboratories Pvt. Ltd,Department of imply primary dengue and < 1.59 secondary dengue infection. DENV 2 was the most
Virology, Haffkine Institute for Training, Research and Testing2, Acharya Donde Marg, commonly associated strain with severe form of dengue in this region.
Parel, Mumbai, Maharashtra, India
Hospital based surveillance of rotavirus associated acute gastroenteritis among
Keywords: AgPath-ID, HiScript, Influenza, One-Step, Real-time PCR under five children from Haryana and Himachal Pradesh after the introduction of
Background: Thermo Fisher Scientific’s AgPath-ID One-Step RT-PCR master mix as the rotavirus vaccine.
approved by Centre for Disease Control (CDC) is used for diagnosis of influenza A
Gupta M1, Bansal A1, Singh M2, Kanojia R3, Muralidharan J4, Bansal A4, Bharti B4, RNA extracted from the culture supernatants detected EV positive by Pan-EV 5’-UTR
Saxena AK5, Sodhi KS5, Kumar R1 assay and further on partial VP1 gene based serotyping were identified as enterovirus
types belonging to EV-A and EV-B species.
Department of Community Medicine and School of Public Health1, Result: A total of 18 different EV serotypes were identified, 3 serotypes such as
PGIMER,Department of Virology2,PGIMER Department of Pediatric Surgery,Advanced Coxsackievirus (CV)-A1, A4,A3 belong to EV-A species , whereas, 15 serotypes such as
Pediatric Centre3PGIMER Advanced Pediatric Centre4PGIMER,Department of Echovirus (E)-2, 3, 5, 6, 11, 12, 20, 25, 30, 33, EVB75,EV-B93, CV-B1, B4, B5 and CV-B6
Radiodiagnosis and Imaging5, Postgraduate Institute of Medical Education and belongs to EV-B species. Among the 18 serotypes CV-A (n=10) and CV-B (n=9) are
Research Chandigarh, India more frequently detected than others, representing 48% of the characterized
isolates.
Email: [email protected] Conclusion: The present study concludes that besides Coxsackie-A and few of
Coxsackie-B viruses, echoviruses
Keywords: Hospital-based, Rotavirus, Acute Gastroenteritis, Surveillance
Background:To monitor trend of rotavirus associated acute gastroenteritis among Prevalence and changing trend of viral markers(HIV, HBsAg and HCV in
under five children after the introduction of rotavirus vaccine in Haryana and hemodialysis patients
Himachal Pradesh, India.
Materials/Methods:Hospital based surveillance system was established in a tertiary Pandey N1, Mittal G2, Ahmad S3, Agarwal KR4
care hospital in Chandigarh, to enrol under five children admitted with acute
gastroenteritis from Haryana and Himachal Pradesh, during September 2016 to May Himalayan Institute of Medical Sciences, Swami Rama Himalayan University
2018, as part of a multicentric study. The rotavirus vaccine was introduced in these
states in 2016. Stool samples (5 ml) were collected, and tested for rotavirus by Keywords – HbsAg , HCV, HIV, ELISA
commercially available ELISA kits (Rotaclone; Meridian Biosciences, USA). Rotavirus Background: Viral hepatitis and human immunodeficiency virus (HIV) infection are
positive specimens were characterized with respect to VP7 (G) and VP4 (P) proteins important causes of mortality and morbidity in patients treated by hemodialysis (HD).
using reverse transcription polymerase chain reaction at Christian Medical College, Hepatitis B virus (HBV) and hepatitis C virus (HCV) are the most important organisms
Vellore. responsible for the patient's morbidity.
Results:Out of.238 children (73.7% males) enrolled (71.8% from Haryana), 236 stool Aims and Objective: To study the prevalence of viral markers (hepatitis B surface
samples were collected. Rotavirus vaccine was administered to 30% of children. antigen , anti HCV and HIV antibody;) in patient undergoing hemodialysis and to
Rotavirus positivity rate was 21.6%. Positivity was similar in both the states. The compare this study with a similar previous study which was conducted in our hospital
highest positivity was in the age group of 0-11 months (20%). The trend of rotavirus 10 years back.
associated acute gastroenteritis was similar in summer (21.8%) and winter seasons Method: All patients( from June 2017 to May 2018) undergoing Hemodialysis at our
(21.4%). G3P[8](18.6%)was the most predominant strain followed by G1P[8](11.8%) center were screened for hepatitis B surface antigen (HBsAg), antibody to HCV (anti-
and G12P[6](9.8%). Fourteen vaccinated rotavirus positive cases, had G3P[8] (4), HCV) and HIV antibody by ELISA and the results were compared with a previous
G1P[8] (2), G1P[4] (2), G12P[8] (2), G12P[6] (1) strains. Vaccine effectiveness was similar study conducted in our hospital 10 years back.
estimated to be 32% after three doses of rotavirus vaccine. Result: A total of 463 patients (both OPD and IPD) were screened for HBsAg, HCV and
Discussion and Conclusions: There was no change in the rotavirus positivity rate HIV infections. Out of 463 patients, 253 were male and 210 were female. 13(2.8%)
(21.6%), as earlier studies have reported positivity rate of 20.9% and 18.8% in this patients were positive for HBsAg, 35(7.6%) for anti HCV and 3(0.6%) for HIV antibody.
region. However, there is change in the most predominant genotype reported i.e., While in the previous study conducted in our hospital, there were 13 patients were
from G1P[8] to G3P[8]. Vaccine effectiveness (32%) was lower than vaccine efficacy HBsAg positive, 36 patients came out to be anti HCV positive and 2 patients were
reported earlier from India (56.3%) and Bangladesh (45.7%). However, these results positive for HIV antibody.
are from one centre, and coverage of vaccination was low (30%). Conclusion: The prevalence of HCV is highest among all the viral markers studied.
Also the prevalence of HBsAg is higher in male patients, that is statistically significant.
Observation and recognition of Non polio Enteroviruses from stool samples of AFP On comparison with the previous similar study, conducted 10 years back, there is a
cases that produced unfamiliar cytopathic effect in L20B cell line statistically significant difference in the prevalence HBsAg and HCV.

Jasmeet Singh1, Harjeet S. Maan1, N.Srivatava1, Tapan Dhole1 and Rachna Chaturvedi2 Machine learning of chemically modified siRNAs and their analysis to advance RNAi
based therapeutics development
Dept. of Microbiology Gandhi Postgraduate Institute of Medical Sciences1, Lucknow,
Uttar Pradesh, India,Amity Institute of Biotechnology, Amity University2, Lucknow. Showkat Ahmad Dar and Manoj Kumar1

Keywords: Acute flaccid paralysis, cytopathic, Intratypic differentiation. Bioinformatics Centre, Institute of Microbial Technology, Council of Scientific and
Background: During laboratory investigation of polioviruses, stool samples producing Industrial Research, Sector 39A, Chandigarh1India
characteristic polio-like cytopathic effect (CPE) in both L20 B cell line and RD cell line
in cell culture are confirmed for polioviruses through Intratypic differentiation-real Email: [email protected]
time polymerase chain reaction (ITD-rRT-PCR). However, few studies only have
shown the susceptibility of L20B cell lines for infection of nonpolio enteroviruses Background: Small interfering RNAs (siRNAs) are extensively used in functional
(NPEV’s). Thus, the present study involves identification of L20B cell grown isolates. genomics and as potential therapeutics. However, they have limitations in delivery,
Method: The cell culture supernatants (n=40) which produced unidentified prone to serum/cellular nucleases, target other similar sequences, etc. that can be
cytopathic effects in L20B cell lines and confirmed as NPEV through ITD-PCR from mitigated by chemical modifications. We have already developed and published
WHO National Polio Laboratory, Department of Microbiology, “siRNAmod an online web resource of chemically modified-siRNAs (CM-siRNAs)”.
Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, Uttar Pradesh, Here, we implemented four methods of machine learning algorithms for model
were analyzed for NPEVs.The culture suspension of these L20Bcell isolates were development. Random forest (RF), Linear regression (LR), Instance based learning
subjected to RNA extraction, panenterovirus 5'-untranslated (UTR) PCR and partial (IBK) and Artificial neural network (ANN) were explored to predict CM-siRNA
VP1 semi-nested PCR followed by sequencing and phylogenetic analysis.The Viral activities. 3031 CM-siRNA dataset with 30 different modifications were used for
model development. Nucleotide (modification) composition, binary pattern features Medicine, London, WC1E7HT, United Kingdom; 5School of Public Health and
and their combinations were executed for 10-fold cross validation and independent Community Medicine, University of New South Wales, Sydney, 2033, Australia.
validation. RF achieved best performance with highest correlation of about 0.85,
while LR, IBK and ANN exhibited 0.66, 0.68 and 0.66 correlation values on training- [email protected]
testing dataset (T2728) respectively. Similar performance was achieved on
independent validation dataset (T303). Subsequently, we analyzed about 150
Background: Molecular techniques for tracing of HIV transmission networks can
modifications on 5000 CM-siRNAs for various chemical fingerprints. This assessment
enhance the power of epidemiological investigations. Identification of the highly
verified their tumorigenicity, mutagenicity, irritant level etc. like properties for their
related clusters by molecular methods could be used to discover sexual and injected
use in medical interventions. We concluded that multiple machine learning based
drug users’ networks. Information about the networks could be particularly useful for
models can be used to predict knockdown activity of CM-siRNAs and further chemical
developing strategies to control HIV transmission among high risk groups (HRG).
fingerprinting will guide for engineering their better versions for therapeutics
Hence, a study was carried out to study transmission patterns among newly
purposes.
diagnosed HIV cases among HRG of North-West India using fingerprinting methods.

Molecular Epidemiology of Influenza Virus Infection in Nepal Methods: Genotyping fingerprinting method was used among 37 randomly selected
samples of recently infected HRGs identified through Recent Infections Testing
Bimlesh Kumar Jha1,A.Sapkota2,S.Dangol2 ,T.Shah2, B. P. Upadhyay1,K.Manandhar1 Algorithm (RITA) using Limiting antigen avidity assay to study transmission patterns
of HIV. Amplification of the reverse transcriptase region of pol (540 base pairs) was
National Public Health Laboratory, Teku, Kathmandu, Nepal; Central Department Of done using Nested PCR. Sequencing was done by ABI DNA sequencer. Reference
Biotechnology,Tribhuvan University,(TU), Nepal sequencing of 11 countries was extracted from HIV Los Alamos database. Alignments
of sequences were done by Clustal X software. HIV-1 subtype was determined on the
*Corresponding Author Email: [email protected] basis of phylogenetic analysis of the pol sequence. Phylogenetic trees were
constructed using the MEGA (version 7.0).
Background::A cross-sectional study in 2018, based at National Influenza Center Result Genotype analysis of 37 recently infected HIV positive individuals revealed 6
Nepal, was carried out with the objectives to isolate and characterize the circulating clusters. One FSW isolate from Punjab and another FSW isolate from Himachal
influenza viruses in Nepal. Pradesh showed linking with Mumbai and Pune isolates. This shows that either FSWs
Material & Methods: A total of 3663 throat swab specimens, obtained from patients or their clients from North- West India move as far as upto Mumbai and Pune for
with Influenza like Illness (ILI) at National Influenza Surveillance Network (NISN) work. The FSW isolates of Chandigarh, Haryana and Punjab are closely related to each
sentinel hospitals, were transported to National Influenza Center, maintaining other showing high movement of FSWs across these states. In men who have sex with
reverse cold chain, within 48 hours. Viral RNA was extracted using QIAmp viral RNA men (MSM) two major clusters were observed. Almost all MSMs from this region
kit. Polymerase Chain Reaction assay (PCR) was performed following CDC Real-time belong to the same cluster showing close networking of MSMs across these states.
rRTPCR protocol for detection and characterization of the influenza viruses including One MSM cluster shared the same clonal clade with Pune isolate showed that the
pandemic influenza virus A (H1N1) pdm 09. Randomly selected 10% of PCR positive MSMs move across the country. Whereas IDU isolates shared the same clonal cluster
specimens were subjected to virus isolation in Madian Darby Canine Kidney (MDCK) with IDU isolates of Chandigarh, Punjab, Haryana and Himachal Pradesh only. No
cells and characterized by Haemagglutination Inhibition Assay.. geographic clustering was observed from North-West and South India. None of the
Results: Out of the 3663 throat swab specimens collected from ILI cases, influenza isolates showed interlinking with other country isolate. This shows that IDUs clusters
viruses were detected in 1206 (39.93%) specimens. InfluenzaA infection was detected are much closed.
in 1004/3663 (11.4%) cases; of which 948/3663 (25.8%) were influenza A (H1N1) pdm Conclusion
09 and 56/3663 (1.5 %) were influenza A/H3 subtype. Influenza B was detected in
Molecular epidemiology methods were able to reveal transmission networks, hence
56/3663 (1.9%) cases. Influenza A (H1N1) pdm09 and influenza B co-infection was
these phylogenetic methods can be used to identify individuals who are within highly
observed in 4/3663 (0.1%) cases. Influenza A (H1N1) pdm 09, A/H3 and B virus were
related transmission groups. Molecular epidemiological methods can be employed to
antigenically similar to the novel influenza A/California/07/2009-Like (H1N1)v type
on a sample of HIV Sentinel Surveillance to monitor transmission networks and
viruses; A/Victoria/361/2011 (H3N2) viruses and B/Wisconsin/1/2010 viruses
primary drug resistance.
respectively. Although sporadic cases of influenza were observed throughout the
year, peak was observed during July to November. The highest number of influenza
Association of FUT2 gene with rotavirus infection among children aged 0-5 years: A
A (H1N1) pdm09 and Influenza B were found in September and in children (<15 years
study protocol
of age group).
Conclusion:All types of influenza viruses are in circulation in Nepal, with the peak
Kaur P1*, Gupta M2, Singh MP3, Sagar V4, Bansal A5, Kanojia R6
during July-November. Comparison of genetic patterns of influenza virus in
consecutive years is necessary to link viral genetic changes with antigenic changes.
Research Scholar, Department of Community Medicine and School of Public Health1;
KEYWORD: Influenza virus, Serodiagnosis, molecular characterization
Professor,Department of Community Medicine and School of Public
Health2,Professor, Department of virology3, Assistant Professor, Department of
HIV transmission patterns study by using genotyping fingerprinting method among
Community Medicine and School of Public Health4,Professor, Advance Pediatrics
the High Risk Groups of North-West India.
Center5,Professor, Department of Pediatrics Surgery6, PGIMER, Chandigarh, India
C.K. Chauhan,1 P.V.M. Lakshmi,1 V. Sagar,1 A. Sharma,2 S.K. Arora,3 R. Kumar1,4,5
Email:[email protected]
1Department of Community Medicine and School of Public Health, Post-Graduate
Keywords: Rotavirus infection, HBGA genotyping, FUT2, secretors, genetic
Institute of Medical Education and Research, Chandigarh, 160012, India; 2Department
susceptibility.
of Internal Medicine, Post-Graduate Institute of Medical Education and Research,
Background:The current understanding of the initial phase of rotavirus acute
Chandigarh, 160012, India; 3Department of Immunopathology, Post-Graduate
gastroenteritis (AGE) and the viral receptor is limited. Rotavirus strain specific
Institute of Medical Education and Research, Chandigarh, 160012, India; 4Department
association with (α-1,2-fucosyltransferase (FUT2) suggested that histo blood group
of Epidemiology and Population Health,The London School of Hygiene and Tropical
antigen (HBGA) serves as an attachment factors for rotavirus. A study protocol to
estimate the prevalence of secretor status and Lewis phenotype; and association of First outbreak of Chikungunya infection in Chandigarh, north India
HBGA with rotavirus infection among under 5 children in North India is presented
here. Chhikara K, Sivanantham K, Binod Pati, Kapil Goyal, Mini P Singh, R.K. Ratho
Methods: A cross-sectional study will be conducted among 360 under five healthy
children in the field practice area of department of Community Medicine, PGIMER, to Post Graduate Institute of Medical Education and Research, Chandigarh,India
estimate the prevalence secretor status and Lewis phenotype. A test negative case-
control study among 242 under five children admitted with AGE with equal number Introduction:
of controls in in a tertiary hospital will be conducted to estimate the association of
Chikungunya is a re-emerging infection in India since 2005 which is caused by
HBGA with rotavirus infection. Rotavirus vaccination status will be obtained to find
Chikungunya virus and of public health importance with the outbreak potential as it
the relationship between susceptibility to infection and its impact on vaccine efficacy.
is transmitted by Aedes mosquito vectors which are more prevalent in India and
Stool (10-20g), and saliva (1-2ml) samples will be collected from enrolled children in
adjacent Asian countries.
both the studies. Secretor status/HBGA and Lewis phenotype will be estimated from
saliva samples using specific monoclonal antibodies. Fecal samples will be tested for
Methods:
rotavirus VP6 antigen using a commercial enzyme immunoassay Rotaclone (Meridian
Bioscience) kits. Rotavirus strains will be characterized by genotyping using reverse The suspected cases with acute onset fever or arthralgia with or without other viral
transcription polymerase chain reaction. Odds ratio of having secretor status among prodromal features during the period of 28th of August 2016 to 25th of February 2017
rotavirus positive AGE cases as compared with rotavirus negative AGE controls, will from 10 states of north India around our PGIMER, Chandigarh were evaluated using
be calculated. Estimation of odds ratio of secretor status among rotavirus serum samples in our virology laboratory for Chikungunya virus infection by IgM
positive/negative vaccinated group as compared with rotavirus positive/negative capture ELISA Kit (NIV Pune) specific for Chikungunya. The sero-positive cases were
unvaccinated group will be done to comment upon the rotavirus vaccine plotted in map manually for geographical distribution.
effectiveness.
Discussion:The results of the study may provide evidence regarding host genetic Results:
susceptibility to rotavirus infection, and its role in vaccine effectiveness.
A total of 1748 serum samples received from Chikungunya suspected patients. A total
Designing a multi-epitope based vaccine to combat Human Cytomegalovirus of 50.34% (880/1748) of suspected cases were positive for Chikungunya IgM capture
(HCMV) infection: An immunoinformatics analysis ELISA. The clinical profile of available data (553/880) from Chikungunya IgM positive
patients was analyzed. The maximum cases reported in the months of October and
Varun Chauhan*, Mini P Singh, Kapil Goyal November and maximum case from Chandigarh, Punjab, Haryana (62%, 22%, 10% of
cases respectively). The cases were involved in all age group with maximum cases
Department of Virology, Post Graduate Institute of Medical Education and Research, were distributed through 3rd to 6th decades of life (13%, 22%, 28%, 17%, 10% of cases
Chandigarh, 160012 respectively) with female (69%) preponderance of 2.23:1 (F:M = 382:171 cases). The
most common clinical feature associated with the cases are fever (97.8%), bleeding
Corresponding Author mail: [email protected] manifestation (46.68%), arthralgia (42.49%), head ache (9.4%), rash (3.2%), myalgia
(2.7%) and thrombocytopenia (1.6%). Data regarding duration of illness and mortality
Introduction: Due to the emerging evidence of drug resistance in HCMV, there is was not available. The maximum sero-positive cases were distributed around the
suppressing need for the development of an effective vaccine. The present study was water bodies, area with migrant and overcrowded population and construction areas
aimed to design a multi-epitope based vaccine targeting the pentavalent complex - of Chandigarh.
gH/gL/UL128-UL130-UL131 of the virus which facilitates virus entry into the host.
Methods: After the application of rigorous immunoinformatics analysis and several Conclusion:
immune filters, the multi-epitope vaccine was constructed, consisting of CD4, CD8
and B cell epitopes. The adjuvant was also linked to the epitopes in order to enhance This is the largest outbreak of Chikungunya in India as for our knowledge with the
the immunogenicity of the vaccine; all were linked via suitable linkers for effective maximum cases in a restricted geographical region with a short span of time which
separation in the host. Since, CMV is a double stranded DNA virus, TLR-9 will primarily emphasis the need of further surveillance and outbreak investigation.
recognise the structural components of CMV. Thus, CpG motifs were added in the
vaccine construct which served as an adjuvant for triggering TLR-9 response. Further,
the binding affinity and stability of the vaccine with Toll like receptor -9 (TLR-9) was Role of Herpes viruses as a causative agent of pneumonia in adult patient with
analysed by molecular docking and dynamics simulation studies. In addition, an in Haematological disorder
silico cloning was performed to ensure the expression and translation efficiency of
the vaccine, utilizing pET-28a (+) vector. Rungta T1, Singh MP1, Khadwal AR2, Chaudhary P1, Kumar A1, Aggarwal R3, Goyal K1,
Results: The proposed vaccine was found to be antigenic and non- allergenic, thus Ratho RK1.
ensuring its immunogenicity and safety. In addition, the epitopes included in the final
Departments of Virology1, Internal Medicine2, Pulmonary Medicine3, Post Graduate
vaccine construct were highly promiscuous thus could have high population
Institute of Medical Education and Research, Chandigarh, India.
coverage. Further, the molecular docking and molecular dynamics analysis indicated
the high affinity of the vaccine with TLR-9. The in silico cloning revealed the
Introduction: Viral infections remain one of the most frequent complications in
acceptable Codon Adaptation Index (CAI) and the acceptable GC content of the
patients with haematological malignancies; majority of these occur due to
optimized nucleotide sequence of the multi-epitope vaccine, thus indicating the
reactivation of latent infection. Among these infections, pneumonia due to viruses
possibility of efficient expression of the vaccine in the host (E. coli -strain K12).
can cause significant morbidity and mortality. The aim of the present study was to
Conclusion: Such T & B-cell-based immunotherapies which leverage this mechanism
find out the prevalence of pneumonia due to herpes viruses (CMV, HSV & VZV) in
could prove their potential against HCMV infection. Further, the authors propose to
patients with haematological disorders undergoing
test the present findings in the lab settings to ensure the safety, immunogenicity and
chemotherapy/immunosuppressive therapy.
efficacy of the presented vaccine which may help in controlling the infection.
 The high sero-prevalence of rubella among pregnant women in absence of childhood
immunization indicates continued transmission of rubella virus. However,
Methods: A total of 29 blood and bronchoalveolar lavage (BAL) samples were considering that nearly 14.3% of the women are still sero-negative, the introduction
collected from patients (Mean age: 44.2, with M:F ratio: 2.2:1) with haematological of rubella vaccine will be useful in the Indian scenario and will help to further lower
malignancies with suspected viral pneumonia prior to the initiation of antiviral the incidence of CRS in the country.
treatment. These were tested by conventional and Real Time (IVD approved) PCR
targeting the immediate early gene of CMV (158bp), glycoprotein D region of HSV Measles and Rubella Outbreaks in North India: 3 year trend
(272bp) and ORF 28 region of VZV (355bp).
K Sangeetha*, Sharma M*, Sharma V*, Rana V*, OP Kumar*, Singh MP*, Goyal K*,
Results: Out of 29 samples, the CMV and HSV were found positive in 10/29 (34.4%) Ratho RK*, Sangal L**
and 4/29 (13.7%) in BAL samples while 3/29 (10.3%) and 2/29 (6.8%) respectively in
blood samples by conventional PCR. Similarly, by Real Time PCR, the CMV and HSV *Departments of Virology, Post Graduate Institute of Medical Educationand
were found positive in 14/28 (50%) and 4/9 (44.4%) in BAL samples while 4/25 (16%) Research, Chandigarh, India and **NPSP WHO, New Delhi
and 2/9 (22.2%) respectively in blood samples. All the samples were negative for VZV
by both the techniques. Imaging findings suggestive of viral pneumonia were seen in Corresponding author email: [email protected]
92.3% (24/26) patients which included consolidation (30.7%), effusion (19.2%), and
nodules (42.3%). The underlying diagnosis was AML (6/29, 20.6%), ALL (5/29, 17.2%), Introduction
NHL and haemolytic anaemia in four (4/29, 13.7%) patients each, multiple myeloma Measles and rubella are highly contagious acute exanthematous febrile
(3/29, 10.3%), T-cell lymphoma, hairy cell Leukaemia in two (2/29, 6.8%) CLL, Post illnesses, commonly occurring as outbreaks. Both these are vaccine preventable
transplantation and myelodysplastic syndrome one each (1/29, 3.4%). diseases. The 11 Member States of WHO SEA-Region committed to eliminate measles
and control CRS by 2020. The six countries to successfully eliminate measles in our
Conclusion: Viral pneumonia due to herpes viruses is a serious threat for region are Bangladesh, Bhutan, Maldives, Nepal, Sri Lanka and Timor-Leste
haematological malignancies. Due to similar clinical picture it becomes important to This epidemiological study was conducted to analyse the trend of measles
differentiate between HSV and CMV so far as the treatment is concerned. Physicians and rubella outbreaks in North India over a period of three years.
dealing with haematological malignancies should be aware of the prevalence and the Materials and Methods
causative virus in order to initiate specific antivirals in patients with high degree of Data from March 2015 - March 2018 was collected through on-going
clinical suspicion. outbreak-based measles rubella surveillance project of MoHFW under the technical
guidance of WHO. Between February 2018 and March 2018, samples of case-based
Sero-prevalence of rubella among pregnant women in a tertiary care hospital, surveillance were also received. The surveillance areas consisted of Chandigarh and
North India its neighbouring states. Representative samples (usually five) were received in cold
chain and samples were tested as per WHO testing algorithm using commercial
Urvashi Nehraa, Sanjay Vermaa, Mini P. Singhb, R.K. Rathob, Ravinder Kaur Sachdevaa,
Siemens Enzygnost kits.
GRV Prasadc, Vanita Suric
Results
Department of Pediatricsa, Virologyb and Obstetricsc, Postgraduate Institute of A total of 471 serum samples were received for testing out of which 221
Medical Education and Research, Chandigarh (46.9%) were positive for measles. Out of 248 measles negative samples tested for
rubella, 96 (38.7%) were positive. Measles positivity was higher [72.5% (95/131)] than
Introduction
rubella [44.1% (15/34)] in 2015. However, the positivity reversed in 2016 with higher
Rubella is an acute contagious viral infection of children which is self limiting in rubella positivity [65% (69/106)] than measles [17.8% (23/129)]. The trend again
nature. However, it is of concern when the infection occurs especially in the first reversed in 2017 with higher measles positivity [35.7% (44/123)] than rubella [13.9
trimester of pregnancy when the child can be born with congenital rubella syndrome (11/79)] and continued the same for the three months of 2018. Throughout the years,
(CRS).Successful vaccination and surveillance helped several countries to eliminate maximum number of outbreaks and positivity was observed during April to June.
rubella/CRS. India introduced measles-rubella (MR) vaccine in 2017, and targeting Children aged 4-9 years contributed to highest positivity for both measles (44.7%)
children aged 9 months to 14 years. An important tool for the evaluation of and rubella (51%). The number of laboratory confirmed outbreaks decreased
vaccination programs is the serological surveillance among pregnant women which consecutively through all three years.
provides information about population immunity profiles and identifies high-risk Conclusion
population subgroups. There is a cycling trend of Measles and rubella outbreaks in North India
during 2015-2018. Due to the introduction of MR vaccine, a reduction in the number
Methodology of outbreaks was observed. The maximum contribution was by children aged 4-9
The present study was conducted among 300 pregnant women in the age group of years, however, increased sero-positivity was observed for 9-14 yrs age group. This
19-45 years attending antenatal clinics in a tertiary care hospital. A maximum of 10 indicates the importance of introduction of second booster dose of vaccination. The
pregnant women were enrolled per day in order to maintain quality of data recent shift from outbreak-based to case-based surveillance should be continued for
collection. Approximately 2 ml blood sample was collected from each pregnant marching towards the goal to eliminate measles and control CRS by 2020.
woman and tested for the presence of rubella IgG antibodies using commercial ELISA
kit.
Cytokine Profiling In Dengue Patients And Its Correlation With Dengue Severity
Results

Out of 300 pregnant women tested, 43 (14.3%) were found to be sero-negative. The Patil S1, Ravi V2, Desai A3
majority of the sero-negative women were in the age group of 26-30 years. Nearly
1,2 & 3
4.65% women were found to be illiterate and 53.48% women were residing in rural Dept of Neurovirology, National Institute of Mental Health and Neurosciences,
areas. Bangalore.

Conclusion Email address: [email protected]

 DENV-1/DENV-4, 5 with DENV-2/DENV-3, and 5 samples with DENV-3/DENV-4. 16
Keywords: Dengue, Cytokine assay, IL10 samples showed infection with 3 serotypes.
Background: Dengue fever is one of the most important mosquito borne viral
infections. India is considered as hyper-endemic country for dengue. Dengue has Conclusions: The present study provides important information on genetic
varied manifestations ranging from subclinical infection to fatal shock syndrome. This diversity of the circulating dengue virus in Mangaluru, Karnataka, India.
study aimed to profile serum cytokine levels in dengue patients and correlate them Concurrent infection with dual and triple serotypes and its co-relation with the
with severity of dengue. severity was observed during the outbreak. The need for continuous monitoring
Material and methods: We conducted cross sectional case control study of 50 of genetic diversity among dengue virus is thus important and will be useful to
serologically confirmed (NS1 Antigen/ IgM antibody positive) dengue cases and 25 study disease burden.
healthy controls over a period of one year. Serum levels of a total of 17 cytokines (IFN
α, IFN γ, TGF β, IL 10, IL1 β, IL 6, IL2, IL4, IL13, IL12p70, IL17 A, VEGF, GMCSF, MCP-1,
Keywords: Dengue virus, dengue, serotypes, genotypes, RT-PCR
TNF α, TNFR1, TNFR2) were analysed using cytometric bead array in cases as well as
controls.
Results: Clinical evaluation of enrolled 50 cases revealed, 41 cases of dengue fever, Prevalence of viral agents among cases of gastroenteritis in a tertiary hospital in
06 cases dengue fever with warning signs & 03 cases of severe dengue. Amongst 50 northern India.
cases, NS1 antigen positivity noted in 40%, IgM positivity in 2%, both NS1 antigen and
IgM antibody positivity noted in 58% cases. Out of the 17 cytokines analysed, mean Ujjala Ghoshal1, Shikha Verma1, Juhi Sisodia1, Nidhi Tejan1, Uday C Ghoshal2
levels of 12 cytokines (IFN α, IFN γ, TGF β, IL 10, IL1 β, IL 6, IL2, VEGF, MCP-1, TNF α,
TNFR1, and TNFR2) were elevated in patients compared to controls. However, the Department of Microbiology1,Department of Gastroenterology2, Sanjay Gandhi
levels of 5 cytokines (IL10, IFNα, IFNγ, MCP-1 & TNFR2) were significantly higher Post Graduate Institute of Medical Sciences, Lucknow
compared to controls (p value 0.05). The levels of IL10 were significantly elevated (p
value 0.015) in severe dengue as compared to dengue fever.
Introduction: Routinely, most of the laboratories test for bacterial and parasitic
Conclusion: Levels of IL10, IFNα, IFNγ, MCP-1 & TNFR2 cytokines are significantly
agents for diarrhoea. A high percentage of cases are clinically suspected to be
elevated in dengue patients compared to healthy controls. The levels of anti-
of viral etiology. Multiplex PCR is a highly sensitive method for detecting the
inflammatory cytokine IL 10 is significantly elevated in severe dengue and thus maybe
enteropathogens. Missing these leads to inappropriate patient treatment and
considered as predictor of severe dengue.
is detrimental to infection control. Recently, few real time PCR based panels
have been introduced. This study was planned to look for viral agents in
Detection of multiple serotypes of dengue virus in acute febrile illness from diarrhoeal stool samples using Fast Track diagnostics gastroenteritis kit.
Karnataka, India

Objective: To determine the prevalence of viral gastroenteritis by testing fecal

Deepika UR1, Rai P*1, Deekshit VK1, Kumar BK1, Chakraborty A1, and Karunasagar I1 samples from patients presenting with diarrhoea.

Email: [email protected] Material and methods: Patients suffering from gastroenteritis were included in
the study from August 2017 till June 2018. Demographic and clinical details of
1Nitte
University Centre for Science Education and Research, Paneer campus, Nitte the patients were recorded in a predesigned proforma. Routine microscopy of
(deemed to be University), Deralakatte, Mangaluru, Karnataka, India-575018 saline and iodine mounts, modified kinyoun staining and culture were done to
exclude parasitic and bacterial agents respectively. Multiplex PCR was done on
Background: Dengue, an important arboviral disease has become a major global extracted nucleic acids using a commercially available kit from Fast track
concern. According to the World Health Organization report, over 3.9 billion diagnostics which detects six important viruses namely: Adenovirus, Rotavirus
people are at risk of dengue virus infection in 128 countries, and more than 50- Norovirus GI, GII, Sapovirus and Astrovirus by Taqman technology.
100 million infections occur each year. The disease burden due to dengue infection
is increasing in India, but there has been limited information on the circulating Results: 70 patients suffering from gastroenteritis were included in the study
strains. Therefore, it is important to determine the serotype prevalent and from August 2017 till mid of June 2018. The mean age of patients was 45 years
circulating in a particular geographical location from time to time. The present and male(n=47) to female(n=23) ratio was 1.9:1. Viral pathogens were
study aims to detect the serotypes associated with the cases of acute febrile illness recovered in 17% (12/70) samples by multiplex PCR. All except one patient were
in Mangaluru during the period of 2018 from an urban background.Among patients with viral gastroenteritis, the ratio
of males: females was 2.25:1(9:4).Of these Norovirus GII (58.1%; 7/12) was the
Methods & Materials: Blood samples were collected from suspected dengue cases most prevalent followed by Astrovirus(25%; 3/12), Adenovirus (17.5%; 2/12),
with febrile illness from Mangaluru. They were screened for NS-1 antigen of DENV, Sapovirus(8.3%; 1/12). Rotavirus was not found in any of the samples. A case of
IgM, and IgG (Dengue NS-1 + Ab Combo Kit). RT-PCR was performed to detect the dual infection with Norovirus GII and Adenovirus was also detected. Abdominal
presence of dengue virus in serum samples using published primer pairs targeting pain was present in all cases of Norovirus and Adenovirus
CprM gene. Dengue virus serotypes were further characterized by nucleotide infection(100%).Vomiting was a feature associated with all Astrovirus infections
sequencing. while it was absent in others except a single case of Norovirus infection. Fever
was present in only one patient which was positive for Norovirus GII.

Results: Out of a total of 112 serum samples from suspected cases of dengue
infection, 103 (91.9 %) samples were positive for DENV with a high prevalence of Conclusion: The prevalence rate of viral gastroenteritis is lower than that
DENV-3 in 78(75.7%) samples followed by DENV-1 in 48 samples (42.8%). 57 reported from other parts of country. In this study, the highest prevalence was
(55.33%) samples had co-infection with more than one DENV serotype out of seen for Norovirus infections which is considered to be the second most
which, 20 were infected with DENV-1/DENV-3, one with DENV-1/DENV-2, 11 with common virus associated with acute gastroenteritis. Also, the number of
children were less which may explain the absence of Rotavirus infections.
Plant Virology approximately 15 x 750 nm containing a molecule of single-stranded, positive-sense
RNA of approximately 10,000 nucleotides. The BCMV genome has a long open
Oral Presentation reading frame (ORF) and a small overlapping ORF, known as “pipo”.Upon expression,
a single large polypeptide is processed by three self-encoded proteases (P1, HC-Pro
Biological and molecular characterization of Soybean yellow mottle mosaic virus and NIa-Pro) to yield ten functional proteins: P1, HC-Pro, P3, 6K1, CI, 6K2, NIa-VPg,
soybean isolate from India NIa-Pro, NIb, and CP. Widespread nucleotide variations were detected throughout
the genome, with significantly more variations observed in 5’ UTR region.
Sandra N1, Tripathi A1, Mandal B2 Comparative analysis of the amino acid sequences revealed that the P1, P3 and N-
terminal region of the CP proteins exhibited higher level of variations than other
(Sandra Nagamani: PhD, Scientist; Tripathi A: MSc, JRF; Mandal B: PhD, Principal cistronic regions. Moreover, selection analyses further confirmed that a number of
Scientist) sites within the P1 and P3 genes have suffered positive selection. These obtained
BCMV sequences also exhibit high recombination frequencies, indicating a more
1Divisionof Seed Science and Technology, 2Advanced Centre for Plant Virology, Indian dynamic evolutionary history. Six isolates with 69 other BCMV genomes, Phylogenetic
Agricultural Research Institute, New Delhi-110012, India analysis showed that both soybean-infecting BCMVs (group I) and peanut-infecting
BCMVs (group II) are distantly related to other BCMVs, suggesting ancestral
Email: [email protected] differentiation and host adaptation. Lastly, group III and IV comprised 17 previously
known BCMV sequences, with the strains in this group isolated from various plant
Keywords: Soybean yellow mottle mosaic virus, Host range, Mechanical sap hosts, including common bean, cowpea, mung bean, purple bush bean, peanut and
inoculation, DAC-ELISA. also soybean. Furthermore, group I was divided into three highly supported
Abstract subgroups (I-a, -b, -c and -d), which is generally in accordance with their geographic
Background: Soybean yellow mottle mosaic virus (SYMMV) belongs to the genus origins
Gammacarmovirus with in the family Tombusviridae. Recently SYMMV mungbean
isolate exhibiting mottling and puckering symptoms was reported from India. So, we Sequence variability and recombination analysis of begomoviruses associated with
surveyed the soybean initial varietal trials (IVT) in the experimental fields of IARI, New yellow vein mosaic and enation leaf curl disease of okra in India
Delhi for the presence of SYMMV.
Materials and Methods: Leaf samples collected from soybean IVT lines subjected to Zainul A. Khan, Madhvi Naresh, Indranil Dasgupta
DAC-ELISA with SYMMV polyclonal antisera and positive samples maintained on
French bean cv. Pusa Parvati by mechanical sap inoculation. Total RNA isolated from Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-
the purified virus preparation, amplified with terminal primers, cloned and 110021, India
sequenced. Host range study was conducted through mechanical sap inoculation on [email protected]
plant species belonging to families Cucurbitaceae, Leguminosae and Solanaceae and
infection confirmed by DAC-ELISA and RT-PCR with CP specific primers. Further real Key words: Begomovirus, Bhendi yellow vein mosaic disease, Okra enation leaf curl
time PCR was performed to find out the most and least susceptible plant species. disease, Recombination
Results: Three samples reacted positively in DAC-ELISA with SYMMV polyclonal
antisera. Sequencing showed that SYMMV soybean isolate consist of 3974 nt with six Introduction
ORF’s and 67 nuclotide difference from mungbean isolate. BLAST analysis showed Okra/Bhendi (Abelmoschus esculentus) belonging to the family Malvaceae, is an
SYMMV-Sb is 98% similar to SYMMV-Mb isolate and 76% with SYMMV-Sb isolate of important vegetable crop of India. India is the largest producer of okra in the world.
South Korea and USA. Host range studies revealed that SYMMV-Sb isolate induced Yellow vein mosaic and enation leaf curl disease of okra is the most important
veinal mottling, mild mottling and chlorotic blotching on soybean, mungbean, constraint to its cultivation in India. These diseases are caused by begomoviruses and
blackgram, cowpea and French bean at 10-15 dpi respectively. Real time PCR data associated betasatellites.
showed that Pusa Parvati and Kentucky wonder were most and least susceptible from Methods
French bean, IPU and P713 was the most and least susceptible varieties of Urdbean. Okra plants showing typical symptoms of yellow vein mosaic and enation leaf curl
Conclusions: This study showed that Indian SYMMV-Sb isolate is 35 nt shorter than disease were collected from Maharashtra, Gujarat and Uttar Pradesh, India. Total
Korean isolate and induced severe symptoms compared to Indian mungbean isolate genomic DNA was isolated from symptomatic and non-symptomatic okra plants. Full
and Korean soybean isolate. Further, this is the first report of severe strain of length genome of begomoviruses and associated satellite DNAs were amplified using
SYMMV-Sb from India. rolling circle amplification kit, cloned into pGreen0029 vector and sequenced.
Infectious clones of begomovirus and betasatellite were constructed and used for
Biological and molecular characterization of bean common mosaic virus isolates in agroinoculation of Nicotiana benthamiana and okra plants.
cowpea Results and Conclusions
Sequence analysis of begomovirus isolated from yellow vein mosaic diseased samples
B.S. Pavithra, M. Krishnareddy, Akshatha Gad, S. Jalali and D.K.Samuel shared maximum identity (92%) with Bhendi yellow vein India virus (GU112050),
while the sequence data of begomoviruses isolated from enation leaf curl diseased
Division of plant pathology, ICAR-Indian Institute of Horticultural Research samples showed maximum identity (92% to 99%) with Okra enation leaf curl virus.
The full length sequence of betasatellites revealed maximum identity (99%) with
E-mail: [email protected] Bhendi yellow vein mosaic betasatellites and alphasatellites isolated in this study
shared maximum identity (95%) with Okra leaf curl alphasatellite. Transmission of
Bean common mosaic virus (BCMV), is a species of the genus Potyvirus (family begomovirus to host plants and putative recombination of begomovirus will be
Potyviridae) is an economically important virus affecting the yield and quality of bean discussed.
and cowpea due to its regular occurrence, ubiquitous seed borne nature and wide
legume crop host range. BCMV isolates collected in Anndhra Pradesh, Karnataka and Betasatellite encoded βC1 protein can hydrolyse ATP and regulates pathogenesis
Tamil Nadu showed symptoms of mosaic, vein banding, leaf deformations, blistering,
and caused significant yield losses (50-100 %). BCMV consists of flexuous filaments Gupta Na, Gnanasekaran Pa, Chakraborty Sa
 response to Tungro infection. Further, the RTSV genome was screened for potential
Molecular Virology Laboratory, School of Life Sciences, JNU, New Delhi, India - RNAi suppressor.
110067.
Results:
[email protected] The result of this study shows that the rice RNA silencing machinery is more active
against the DNA virus (RTBV) compared to the RNA virus (RTSV), both the viruses
Key words: being part of the tungro virus complex. Further, RTSV proteins have very weak
ToLCPB, ATPase, βC1 suppressor activity against RNA silencing, suggesting the presence of alternate
Background information: silencing mechanisms. In addition, a large number of rice micro RNAs, which are
In the recent years, role of betasatellites in geminivirus-disease complex has been crucial for the control of gene expression during various plant developmental stages
recognized worldwide. Betasatellites are circular, single stranded DNA molecules are differentially regulated upon tungro infection.
which are associated with the monopartite begomoviruses and intensify the disease
development. Betasatellites encodes a single protein, βC1 that favours pathogenesis Conclusions: This study helps elucidate the elementary framework of the host RNAi
through suppression of several anti-viral defence mechanisms in plant. Many of the defence response against Tungro infection in rice opening avenues for engineering
viral protein shows non ATPase activity. Bioinformatics analysis revealed that βC1 resistance against the virus complex in rice.
possess ATP grasp motif.
Materials and methods:
Protein purification was carried out by affinity chromatography followed by ion Topical application of double-stranded RNA molecules containing sequences of
exchange chromatography. The proteins were detected through western blotting Tomato leaf curl virus and Cucumber mosaic virus provides resistance against the
analysis using anti GST antibody. ATPase assay was done by polyethyleneimine thin cognate viruses.
layer chromatography method. To detect viral DNA level, Southern hybridization, leaf
Namgial T1,2, Kaldis A1, Chakraborty S2, Voloudakis A1,*
disc assay was done.
Results: 1 Laboratory of Plant Breeding and Biometry, Department of Crop Science, Agricultural

In this study, the purification and biochemical characterization of βC1 protein University of Athens, 11855 Athens, Greece
encoded by Tomato leaf curl Patna betasatellite (ToLCPB) was carried out which 2
School of Life Sciences, Jawaharlal Nehru University, 110067 New Delhi, India
exhibited a novel ATPase activity. Amino acid residue Lys-49, Arg-69 and Arg-91 are
important for ATPase activity. The ATPase activity was found to influence the viral [email protected]
DNA and transcripts level. To know whether the ATPase activity is conserved among
betasatellites, comparative assays were carried out. Nicotiana benthamiana plants Keywords CMV ∙ Double-stranded RNA ∙ Geminiviruses ∙ RNA interference ∙ RNAi ∙
infected with different betasatellite alongwith with Tomato leaf curl virus indicated RNA-based vaccination ∙ ToLCGV.
differential infectivity pattern. Introduction
Conclusions: Geminiviruses compose the largest plant-infecting group of ssDNA viruses, they are
The newly identified role of ToLCPB βC1mediated ATP hydrolysis regulates viral transmitted through insect vectors and infect several economically important crops
pathogenesis in plants. Critical role of the amino acids of βC1 protein governing causing high yield losses. Tomato leaf curl virus (ToLCV) is a bipartite geminivirus
ATPase activity have been identified. Comparative efficacy of different betasatellites causing serious damage in tomato in India. Cucumber mosaic virus (CMV) is also a
has been determined for ATPase activity and pathogenesis. serious pathogen of tomato; CMV-infected tomato plants are stunted, bushy and may
have malformed leaves and fruits. RNAi that is triggered by double-stranded RNA
Contextualizing RNAi defence response against virus infection in rice (dsRNA) molecules, is a powerful means to control plant viruses in transgenic plants,
as well as in a non-transgenic manner in a process designated as ‘RNA-based
Fauzia Zarreen and Indranil Dasgupta vaccination’.
Methods
Department of Plant Molecular Biology, University of Delhi South Campus, Benito DsRNA molecules were made for the overlapping regions of ToLCV, namely AC1/AC4,
Jaurez Road, New Delhi-110021 AV1/AV2, AC1/AC4_AV1/AV2 (the latter designated as fusion construct), for CMV-2b
and CMV-2b_ToLCGV-AV1/AV2 (the latter designated as hybrid construct, containing
[email protected] regions from two different viruses).
Result
Key words: RNAi, siRNA, VsiRNA, miRNA, RNAi suppressor In tomato, dsRNAs for AC1/AC4, AV1/AV2, AC1/AC4_AV1/AV2 and CMV-2b_ToLCGV-
AV1/AV2 conferred 45%, 60%, 50% and 55% resistance against ToLCGV, respectively.
Introduction: Experiments with Nicotiana tabacum showed that the dsRNA construct CMV-
Plant viruses trigger strong RNA silencing response in host plants and subsequently, 2b_ToLCGV-AV1/AV2 conferred 33.3% resistance against CMV, while CMV-2b
the viral transcripts are targeted by the virus-derived siRNA which accumulate to high provided 40% resistance.
levels during virus infection. The major class of siRNA produced in response to virus Conclusion
infection is 21-25 nt siRNAs (VsRNA). Besides, virus infection also results in changes The present study also reported that the dsRNA exhibits systemic transport in tomato.
in the host miRNA expression profile. In course of evolution, viruses have developed This is the first case where RNA-based vaccination is functional for a bipartite
strategies to counteract the RNAi-based defence of the plants. Rice Tungro disease is geminivirus and where a single dsRNA molecule could induce resistance against two
an important viral disease of rice affecting the rice crop in the south and south-east tomato-infecting viruses, namely an RNA (CMV) and a DNA (ToLCV) plant virus.
Asia caused by the joint infection of Rice tungro bacilliform virus (RTBV) and Rice Therefore, the production of single dsRNA molecules containing the desired targets
tungro spherical virus (RTSV). could provide control against different viruses infecting the crop of interest.

Methods: Red seaweed-based formulations for effective management of plant viral diseases
With the aim of examining the plausible RNAi defence mounted against Rice Tungro
infection, we analyzed the small RNA (VsRNA and miRNA) produced in rice in Girish TR1, Sumit Bhose1, Nagaraju N2, Sri Sailaja Nori1, Shrikumar S1
 percent transmission (92-100%) in all the independent trials within 21 days post-
1
Sea6 Energy private limited, C-CAMP-NCBS, Bellary Road, Bengaluru, India 560065 inoculation. Confirmed the GBNV infection in the inoculated plants by ELISA and RT-
2Department of Plant Pathology, UAS-GKVK, Bengaluru, India 560065 PCR, and noticed maximum GBNV titer in tobacco seedlings.
CONCLUSION(S): The protocol optimized in this study will be useful in evaluating
[email protected] tomato genotypes for BND resistance and identifying the sources of resistance, and
subsequently in breeding for BND resistance in tomato.
Key words: Natural products, Plant viral diseases, Crop protection KEYWORDS: Tomato, germplasm evaluation, resistance breeding, sap inoculation
protocol, groundnut bud necrosis, orthotospovirus.
Plant viruses pose a serious threat to global agricultural productivity. About 40% of
the total crop loss annually is attributed to several viral diseases. Although the use of Current status of viral diseases of cucurbitaceous crops in India
chemical pesticides has helped in managing plant viral diseases the vector control
alone seems insufficient. Moreover, the indiscriminate use of chemical pesticides has Mahesha, B. and Krishna Reddy, M.
resulted in undesirable consequences on both environment and human health.
Therefore, the combination of vector control methods with other treatment Plant Virology Laboratory, Division of Plant Pathology, Indian Institute of Horticultural
regimens such as boosting up of plant’s innate defense mechanisms using natural Research (IIHR), Hesarghatta Lake Post,
plant defense activators could be more effective and such a treatment regimen may Bangalore-560089.
also help in reducing the usage of chemical pesticides. At Sea6Energy, we aim to
develop seaweed derived molecular solutions that are natural and ecofriendly. Our E-mail: [email protected]
idea is based on the hypothesis that plants possess several layers of innate immunity
that can be activated by certain seaweed derived active ingredients. Based on Abstract
fractional analysis of the Kappaphycus species of cultivated red seaplant biomass, we The field survey was conducted at selective districts of Uttar Pradesh and Karnataka
have identified unique active ingredients that can elicit plant defense pathways. states (including farmer fields, Agricultural regional research stations and
Prophylactic, foliar application of these solutions on various vegetable and experimental plots) at more than 50 villages, to assess the current status of viral
horticulture crops revealed about 20-40% reduction in viral incidences and an diseases of cucurbitaceous crops in and around the region. The major viral diseases
associated overall yield improvement of upto 40%. The transcriptome analysis of observed on cucurbitaceous crops were belongs to the genus Begomovirus (85-
foliar samples treated with these formulations revealed several fold up-regulation of 100%), Potyvirus (30-100%), Cucumovirus (30-80%), Tospovirus (30-100%),
genes related to defense response pathways. Overall, our studies suggest red Polerovirus (30-100%), Tobamovirus (30-90%), Ilarvirus (40-85%) and Crinivirus (30-
seaweed-based formulations can be effectively exploited to manage plant viral 65%) are most predominantly occurring in endemic manner with an incidence of 30-
diseases. 100% causing yield losses (10-60%)and quality (40-80%) loss of the produce.Among
all the viruses Tomato leaf curl New Delhi virus (G: Begomovirus, F: Geminiviridae),
Optimization of mechanical sap transmission method on tomato (Solanum Papaya ring spot virus-w (G: Potyvirus, F: Potyviridae), Cucumber mosaic virus
lycopersicum L.) plants with groundnut bud necrosis orthotospovirus (G:Cucumovirus, F:Bromoviridae), Watermelon bud necrosis virus(G: Tospovirus, F:
Bunyaviridae), Groundnut bud necrosis virus(G: Tospovirus, F: Bunyaviridae), Cucurbit
Basavaraj1*, A. Kumar2, M.K. Yadav1, V. R. Sharma3, B. Mandal1 and R.K. Jain1 aphid borne yellow mosaic virus (G:Polerovirus, F: Luteoviridae), Cucumber green
1Advanced Centre for Plant Virology (ACPV), Division of Plant Pathology, ICAR-Indian mottle mosaic virus(G:Tobamovirus, F:Virgaviridae), Tobacco streak virus(G: Ilarvirus,
Agricultural Research Institute, New Delhi-110 012, India F: Bromoviridae) were predominant in cucurbit ecosystem by causing severe yield
2Division of Vegetable Science, ICAR-Indian Agricultural Research Institute, New and quality loss of the economic produce. The most affected cucurbit crops are
Delhi-110 012, India ridgegourd, pumpkin, bottlegourd, watermelon, cucumber, bittergourd, muskmelon,
3Department of Genetics and Plant Molecular Biology, CSIR-National Botanical squash, spongegourd, snakegourd and ashgourd. All the isolates were collected and
Research Institute, Lucknow, India diagnosed through Transmission Electron Microscope (TEM), Atomic Force
[email protected] Microscope (AFM), DAC-ELISA, PCR and RT-PCR analysis with standard protocols.
Further, developed the virus transmission protocols under artificial conditions and
BACKGROUND: Groundnut bud necrosis orthotospovirus (GBNV) inciting bud identified the host ranges of individual viruses. The mixed infections, contagiousness,
necrosis disease (BND) is a major limiting virus affecting the tomato cultivation in seedborne nature of viruses and insecticide resistance issue, cross infections due to
India and Asia. Evaluation of tomato germplasm(s) for BND resistance and host-virus wide host range leads to severe and rampant spreadof cucurbit viruses. Developed
interactions studies became challenging due to the difficulty in inducing GBNV the Integrated viral disease management strategies in musk melon, cucumber and
infection on tomato. ridgegourd crops that helps the cucurbit growers considerably. Further, ample scope
MATERIAL/METHODS: In this study, a GBNV isolate (from tomato fruit) collected to identify and employ the resistant sources in breeding programs and novel bio-
from the experimental fields and maintained on Vigna unguiculata (cowpea cv. Pusa technological interventions like genome editing (CRISPR/Cas9) techniques can be
Komal), Nicotiana benthamiana (tobacco) and Solanum lycopersicum (tomato cv. explored in the management of cucurbit viruses.
Pusa Ruby) were used as the different sources of inoculum. Performed the sap Key words: Cucurbit viruses, Current status, Diagnosis, Management
transmission on tomato seedlings from these inoculum sources with an inoculation
buffer i.e. sodium phosphate buffer 0.1M (with 0.15% Sodium Sulphite + 0.2% β- Reverse Transcription-Loop Mediated Isothermal Amplification: A rapid diagnostic
mercaptoethanol) prepared at four different pH ranges (6.5, 7.0, 7.5, 8.0). Evaluated assay for detection of Potato virus A in potato and aphids
four different growth stages of tomato seedlings viz., 15, 20, 25 and 30 days after Baswaraj Raigond*1, Verma A1, Pathania S1, Jandrajupalli S2, Verma G1, Kochhar T1 and
sowing (DAS) to ascertain the effective growth stage to achieve maximum rate of Chakrabarti SK1
GBNV transmission. Conducted the transmission experiments in the environment-
controlled containment facility at a temperature range of 26-28oC. 1
ICAR-Central Potato Research Institute, Shimla, Himachal Pradesh-171001, India
RESULTS: Mechanical sap transmission using systemically infected tobacco plants as 2ICAR-National Institute of Biotic Stress Management, Raipur, Chhattisgarh- 493225,
inoculum source resulted in a maximum rate of transmission on tomato when India
inoculated with the buffer of pH 6.5. Most suitable growth stage of tomato seedlings
for GBNV transmission was at 25 DAS, which resulted consistently the maximum [email protected]
 ToLCV symptomatic samples were collected and isolated its genomic DNA. These
Background: Potato virus A is an important virus infecting potato, transmitted samples were tested in PCR with universal primer, Deng A/B and confirmed the
through infected tubers, vegetative planting material and by aphids. It’s one of the presence of begomovirus. However to identify viral strain, designed a specific primer,
major concerns for raising healthy seed potato. Under field conditions, window (TL12F/13R for mono and TL14F/15R for bipartite) using DNA-A genome of ToLCV.
available for monitoring viral incidence, monitoring of viruliferous aphid etc., is The amplified PCR products were sequenced and submitted in NCBI database.
minimal. Hence, draw special attention for developing rapid and sensitive diagnostic
assay to taking-up timely management practices. Hence, we aimed to develop LAMP Results:
assay for specific detection of PVA.
The samples amplified with primer TL12F/13R similar with tomato leaf curl Karnataka
Material/methods: Total RNA isolation and RT-PCR based confirmation of PVA from virus (Acc.No KY311884) and with TL14F/15R similar with Tomato leaf curl New Delhi
pure culture and from experimentally prepared viruliferous aphids was conducted. virus (Acc.No. MH269415). These strains were maintained in greenhouse on tomato
The LAMP assay was optimized and confirmed its specificity and sensitivity. Extended plants through whiteflies transmission. These strains were tested on breeding
for detection in tubers and in single aphids and validated by running across field population with different combinations of Ty genes (Ty-1, Ty-2, Ty-3, ty-5, Ty-1+Ty-2,
collected potato and aphid samples. Ty2+Ty3, Ty2+ty-5, Ty1+Ty3, ty-5+Ty-6 and Ty-1+Ty-2+Ty-3) along with susceptible
control. The healthy whiteflies incubated on tomato plant infested with either mono
Results: A RT-LAMP assay was optimized where, 61oC for 60 min gave sharp ladder or bipartite for 24hrs then released on experimental tomato plants. After infestation,
like amplification and total volume of the reaction mixture was successfully reduced the data was recorded on 20th and 45th day using 0-4 disease scoring. The results after
to 10 μl. Assay was visualized by naked eyes by adding SYBR gold nucleic acid stain. It 45 days against monopartite showed that plants having Ty-2+Ty-3 and Ty-1+Ty-2+Ty-
was found to be highly specific as there was no cross reaction with other viruses 3 were tolerant. However, against bipartite begomovirus, plants having Ty2+Ty3 and
infecting potato. Sensitivity of the assay was equivalent to RT-PCR, were it was able Ty-1+Ty-2+Ty-3 were moderately tolerant after 20 dpi and became susceptible after
to detect the virus at 10-2 dilution. The assay can be successfully applied for detection 45 dpi.
of PVA in potato tubers. Squash print RT-LAMP assay was developed and found
suitable to determine viruliferous nature of aphids. Finally, the optimized RT-LAMP Conclusions: The Ty genes interrogation into commercial tomato lines is useful to
and SP-RT-LAMP assays were successfully validated by running across potato samples alleviate losses due to ToLCV.
and aphid vectors collected randomly from fields respectively.
Understanding the role of glycine rich RNA binding proteins in viral pathogenesis.
Conclusions: Developed RT-LAMP and SP-RT-LAMP assay for specific and rapid
detection of PVA in potato plants and in single aphids respectively which can assist in [email protected]
raising healthy seed potato.
Key words: Potato virus A, Loop mediated Isothermal amplification, detection, Squash Background information: Glycine rich proteins (GRP) represent a group of proteins,
Print, aphid, potato. which are characterized by the conserved semi-repetitive glycine rich motif at its C-
terminal. GRPs are involved in growth, development and other multiple cellular
A comprehensive study on resistance to Tomato leaf curl virus against various processes. GRPs are also induced during various abiotic and biotic stresses but the
combinations of Ty genes in tomato mechanism during biotic stress especially during viral pathogenesis is poorly
observed. Structurally, the Class IV GRPs essentially consist of two domains, a
Pardhasaradhi, P1,2., Rakesh Kumar*1, Sairam, P1., Ramachandran, E1., Santanu structured N-terminus containing an RNA Recognition Motif (RRM) and a flexible C-
Acharya1 and Rajashaker, P2 terminus with numerous glycine making the part intrinsically disordered. Although
key reports are available about the role of Class IV GRPs in biotic stress responses, a
1 JK Agri Genetics Ltd, Hyderabad; 2 JNTU, Hyderabad, role of these proteins in the life cycle of viruses has not yet been intensively studied.
Materials and methods: NbGRP3 transcript level in viral infected samples quantified
[email protected] by northern hybridization/qRT-PCR. NbGRP3 cloned and purified to study the
interaction with viral DNA using EMSA/fluorescence binding.
Key words: Tomato, Ty gene, Begomovirus, Resistance Results: NbGRP3 (a Class IVa GRP from N. benthamiana, a model organism for plant-
virus interaction studies) shows elevated transcript levels upon infection of
Geminiviruses, i.e., Tomato leaf curl New Delhi virus (ToLCNDV) along with non-
cognate Radish leaf curl betasatellite (RaLCβ) in N. benthamiana. Similar kinds of
observations were obtained with Tomato bushy stunt virus (TBSV), which is an RNA
virus. Upon further investigation, it was found that NbGRP3 interacts with Satellite
Conserved Region (SCR) of Radish Leaf curl virus Beta satellite.
Conclusions: Up-regulation of transcript level of NbGRP3 upon viral infection (both
DNA and RNA viruses) and their interaction with viral DNA stress on a role of NbGRP3
during viral pathogenesis. Yet further experiments need to be performed to
understand the precise role of NbGRP3 in viral pathogenesis.
Keywords: NbGRP3 – ToLCNDV – RaLCβ – SCR - TBSV
Introduction: Tomato Leaf Curl Virus (ToLCV), a begomovirus, is a major constraint Characterization of begomovirus and associated satellite components causing leaf
for the production of tomato cause up to 100% crop loss. To overcome these curl disease in Alcea rosea L.
problems, researchers identified Ty-genes from wild tomato species. In this study, we
introgressed these genes into commercial tomato lines and tested its efficacy against Kumar Manish, Kumar RV, Chakraborty S.
begomoviruses.
Molecular Virology Lab, School of life sciences, Jawaharlal Nehru University, New
Methods: Delhi-110 067, India.
[email protected] the produced small RNAs (sRNAs) are loaded onto RISC complex. While interacting
with RISC, the passenger strand is degraded while the guide strand binds to the
Keywords: Hollyhock, Recombination, begomovirus. complementary sequences in the target mRNA leading to either cleavage of the
Introduction: mRNA transcript or translation inhibition. RNA dependent RNA polymerases (RDRs)
Hollyhock (Alcea rosea L. family Malvaceae) is an annual ornamental plant grown in are the enzymes in plants which uses primary small-interfering RNA (siRNA) produced
tropical, subtropical and temperate regions of the world. Geminiviruses infect a large by DCLs, to generate longer dsRNA to be processed into secondary siRNA. The
variety of crop plants around the world and cause a new threat to global food secondary siRNA amplifies the signal leading to the systemic silencing. Many plant
security. Earlier studies reported that begomoviruses are known to infect hollyhock hormones are also regulated by a variety of WRKY and other proteins.
plants. In February 2016, Begomovirus-like symptoms were noticed on the hollyhock Material/methods: We have inoculated N. benthamiana plants and collected leaf
plants grown in a garden at the campus of JNU, New Delhi. Since the hollyhock plants samples (21 dpi) to generate sRNA libraries. We have uses next generation
exhibited typical symptoms of yellow vein mosaic, we anticipated the possibility of sequencing (NGS) to identify differentially expressed sRNAs in N. benthamiana
association of begomoviruses in these plants. following Tomato leaf curl virus (ToLCV) infection.
Methods: Results: Several sRNAs were found to be differentially expressed. Among them genes
Rolling circle amplification was performed to clone the associated begomoviral encoding WRKY-55, Cytochrome P450 like TBP, Laccase, Serine/threonine-protein
genomic components from the total genomic DNA isolated from the infected phosphatase and Phospho-pantothenoylcysteine decarboxylase etc. were up-
hollyhock plants and clones were sequenced. The phylogenetic analysis was carried regulated following virus infection in N. benthamiana. In contrast, genes encoding
out by MEGA-X software and the recombination analyses of the cloned begomoviral Photosystem II CP43 chlorophyll apoprotein, Pyridoxal phosphate (PLP)-dependent
genomic components were analyzed using (Recombination detection program) RDP. transferases, Photosystem II CP47 chlorophyll apoprotein, Integrin-linked kinase-
Results: associated serine/threonine phosphatase 2C were down-regulated.
We have identified a new virus species, Hollyhock yellow vein virus: New Delhi and Conclusions: The in-silico analysis suggests ToLCV infection results in differential
isolates of Cotton leaf curl Multan virus, Ludwigia leaf distortion betasatellite, Cotton expression of sRNAs. Among the sRNAs, those regulating transcription factors or
leaf curl Multan alphasatellite and Sida leaf curl virus alphasatellite in association various enzymes might be involved in plant metabolic pathways.
with yellow mosaic disease. Recent studies also suggest that the begomoviruses have Keywords: ToLCV; N. benthamiana; sRNA sequencing; siRNA
been derived through extensive recombination processes. The study suggests that
the occurrence of recombination events between related begomoviruses may lead to Development of chimeric promoters from parareterovirus for strong expression of
the appearance of the new begomovirus species. ectopic gene in plant system
Conclusions:
We identified a new virus species, Hollyhock yellow vein virus: New Delhi along with Dipinte Gupta*& Rajiv Ranjan
the others begomoviral genomic components, infecting the hollyhock plants. The
infectivity of hollyhock yellow vein virus in different host plant needs to be assessed Plant Biotechnology Lab, Department of Botany, Faculty of Science, Dayalbagh
to confirm the agro-economic threat of the begomoviruses. Educational Institute (Deemed University), Dayalbagh, Agra 282005, India

[email protected]
Identification of Brassica yellows virus, a Polerovirus in Radish (Raphanus sativus)
Promoters are specific sequence of nucleotides present upstream of the gene coding
[email protected] region; comprises of multiple cis-acting elements having specific binding for
transcription factors which are involved in initiation and regulation of transcription.
Survey conducted in the vegetable growing regions of Karnataka indicated incidence The basic strength of promoter lies in the notation and arrangement of these cis-
of virus and virus like diseases with symptoms of yellowing, chlorotic lesions, acting elements and their combinational interaction with transcription factor.
interveinal chlorosis and leaf distortion in radish. The symptomatic samples collected Hereby-strong promoters can be developed by redesigning their architecture, which
from field, were washed with RNase-free sterile double distilled water, sap extracted can be exploited for strong expression of ectopic gene. In the present study, eight
and were examined under Electron microscope, which showed the presence of chimeric promoters were synthesized by using the two well-established approaches:
isometric particles of 28-30nm. Total RNA were extracted from symptomatic leaf domain swapping and hybridization. Promoters from Rice Tungro Bacillus Virus
samples and healthy control were used for RT-PCR amplification using Polerovirus (RTBV)and Mirabilis Mosaic Virus(MMV12) were used as parents for the development
specific generic primers. The results have shown amplification of 350bp DNA of these chimeric promoters. All the developed promoters were cloned upstream of
fragment from infected samples but not from healthy control plant samples. Cloning, aGUS reporter gene and their efficacy was tested by transient as well as transgenic
sequencing and nucleotide blast search showed 93 % similarity with poleroviruses assay. Gus assay of electroporated tobacco protoplast and agro-infilterated tobacco,
and more than 93 to 99% with Brassica yellows virus (BrYV) (Genus: Poleroviruses; petunia, rice and millets was performed for transient expression studies. Transgenic
Family: Luteoviridae). This is the first report of the occurrence of BrYV in radish in lines of tobacco were generated at T0 stage for the expression profiling of developed
India. BrYV has a wider host range and is an emerging viral disease with potential promoters. By doing the expression profiling on biochemical, histo-chemical and
threat. Further complete genome sequence is under progress. molecular level, twoout of the eight developed promoters,designated as REM and
RTBVP1MMV12 were found to have great efficiency. On comparing these developed
Identification of differentially expressed small RNAs following Tomato leaf curl virus promoters with the most widely used promoter CaMV 35S, 18.12% and 32.8%
infection in increase in expression was found in REM and RTBVP1MMV12 respectively in dicots
while in monocots 30% and 98.64% increasewas found in REM and RTBVP1MMV12
[email protected] respectively. From the obtained results it might be concluded that we developed
Background Information: The phenomenon of RNA silencing protects host from promoter can be efficiently used for translational research in plants and can pave the
viruses and in addition can also silence endogenous heterochromatin and way off for plant biotech industries.
transposons, transcriptionally or post-transcriptionally in a sequence-specific
manner1, 2. The trigger for this process are double stranded RNA (dsRNA) precursors. Development of efficient synthetic Promoter from plant parareterovirus
DICER-LIKE (DCLs) proteins in plants, which are RNase III-type endoribonucleases,
cleave dsRNA into short RNA duplex of 21-24 nucleotide in length3, 4. Subsequently, Dipinte Gupta*& Rajiv Ranjan
Plant Biotechnology Lab, Department of Botany, Faculty of Science,
Dayalbagh Educational Institute (Deemed University), Dayalbagh, Agra 282005, India Conclusions: Differential responses of CMV severity might be due to vector load,
[email protected] climatic conditions and genetic characteristics of chilli lines. These highly resistant
and resistant lines could be used by breeders in developing new chilli hybrid resistant
Promoters are specific sequence of nucleotides present upstream of the gene coding to CMV.
region; comprises of multiple cis-acting elements having specific binding for
transcription factors which are involved in initiation and regulation of transcription. Emerging virus and phytoplasma diseases in Rajouri District of Jammu and Kashmir
The basic strength of promoter lies in the notation and arrangement of these cis-
acting elements and their combinational interaction with transcription factor. Mohd Ashaq
Hereby-strong promoters can be developed by redesigning their architecture, which
can be exploited for strong expression of ectopic gene. In the present study, eight Higher Education Department, J&K Govt
chimeric promoters were synthesized by using the two well-established approaches:
domain swapping and hybridization. Promoters from Rice Tungro Bacillus Virus email id: [email protected]
(RTBV) and Mirabilis Mosaic Virus(MMV12) were used as parents for the
development of these chimeric promoters. All the developed promoters were cloned Key words: Survey, Virus, Phytoplasma, Rajouri District, Jammu and Kashmir,
upstream of aGUS reporter gene and their efficacy was tested by transient as well as Identification, different symptoms, Climate change.
transgenic assay. Gus assay of electroporated tobacco protoplast and agro-
infilterated tobacco, petunia, rice and millets was performed for transient expression
studies. Transgenic lines of tobacco were generated at T0 stage for the expression Introduction:
profiling of developed promoters. By doing the expression profiling on biochemical,
histo-chemical and molecular level, twoout of the eight developed Climate change has triggered a number of plant viral diseases in recent past which
promoters,designated as REM and RTBVP1MMV12 were found to have great were never observed or recorded before. There is urgent need to detect these newly
efficiency. On comparing these developed promoters with the most widely used emerging diseases and provide control measures to ensure the better plant heath
promoter CaMV 35S, 18.12% and 32.8% increase in expression was found in REM and hence greater food security. In this connection, a random survey was conducted to
RTBVP1MMV12 respectively in dicots while in monocots 30% and 98.64% study the virus and phytoplasma infecting important vegetables, fruit plants,
increasewas found in REM and RTBVP1MMV12 respectively. From the obtained ornamentals and weeds in District Rajouri of Jammu and Kashmir State (India).
results it might be concluded that we developed promoter can be efficiently used for Vegetables like potato, chili, turnip, radish, cabbage, cauliflower, knoll kohl, peas,
translational research in plants and can pave the way off for plant biotech industries. okra, brinjal, and cucurbits (bottle gourd, cucumber, snake gourd and bitter gourd
etc), wild fruiting plants like figs and blackberries, ornamentals (wild rose, marigold,
Identification of chilli (Capsicum annuum L.) lines with enhanced resistance to Catharanthus etc.) and weed hosts were surveyed. In many plants, severe effect on
cucumber mosaic virus under controlled and field conditions plant health and crop losses due to virus and phytoplasma infections was observed.
Several virus infected samples were collected for further biological and molecular-
Kavyashri V Va, Nagaraju N b and Mohan Rao A c based investigation. Different kinds of virus or virus-like symptoms like mosaics, vein
clearing, vein banding, ring spot, leaf curling, leaf rolling etc in different crops/plants
email: [email protected] were observed. During survey, in many cases, different kinds of symptoms in infected
plants were observed, which were not recorded previously, it might be due to the
Key words: Screening, chilli, CMV, Disease Incidence (DI), ELISA. effect of climate change that influences the pathogens to become aggressive and
more virulence or forcing to shift to the new hosts.
Introduction: Cucumber mosaic cucumovirus (CMV) is a destructive pathogen with a
wide host range. Identification of resistant sources and understanding the genetic Methods: Initial field surveys have been conducted and samples are collected for
behavior of resistance is essential for resistance breeding. further investigation. Diagnostic techniques for virus identification include 1.
biological properties related to the interaction of the virus with its host and/or the
Methods: One hundred and eleven chilli lines collected from public and private vector (symptomatology and transmission tests) and 2. intrinsic properties of the
institutions were evaluated for CMV resistance both under field and greenhouse virus itself (coat protein and nucleic acid). Detection methods based on coat protein
conditions. Reaction of chilli lines for CMV was visualized through symptoms and include precipitation/agglutination tests, enzyme-linked immunosorbent assays, and
confirmed by Double Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS- immunoblotting whereas viral nucleic acid-based techniques are like dot-blot
ELISA). hybridization assays and polymerase chain reaction (PCR).

Results: The CMV disease incidence (DI) was ranging from 0.0-100%, six chilli lines Results: The results are awaited as initial survey has been completed and molecular
viz., AVPP 0906, AVPP 1110, Aparna, Susanjoy, Phule Jyothi and ADR Driver were detection is going on.
remained free from CMV infection with least disease progress (AUDPC) therefore
catalogued as highly resistant. Rest of the lines exhibited characteristic CMV Conclusions: The survey of virus and phytoplasma diseases on virus various plant
symptoms. Twelve lines namely, AVPP 0302, AVPP 0508, AVPP 1111, A. Khyati, Assam species provide a clue on the impact of climate change on plant health may open new
2, HMT 1, Byadagi dabbi, Ujwala, LCA 960, AVPP 0904, Wakako long and Utkal Rashmi vistas for investigation and controlling plant diseases.
were found resistant and moderately resistant reaction both under field and
greenhouse conditions. However under mechanical inoculation, six lines were First report of Soybean Yellow Mottle Mosaic Virus on Glycine max in India.
categorized as highly resistant, nine as resistant, three as moderately resistant, 19 as
moderately susceptible and 74 as susceptible. Whereas under field condition, six lines [email protected]
showed highly resistant reaction to CMV, five lines showed resistant reaction, eight
lines were moderately resistant, three lines were found moderately susceptible, Soybean yellow mottle mosiac virus (SYMMV) belongs to the genus Carmovirus with
ninety one lines were found susceptible to CMV based on DI. in the family Tombusviridae has been recently reported from mungbean in India.
Later, while surveying for other leguminous hosts of SYMMV in the fields of Indian
Agricultural Research Institute, New Delhi, three soybean samples strongly reacted Advanced Centre for Plant Virology, Division of Plant Pathology, ICAR-Indian
with the polyclonal antibodies raised against SYMMV in direct antigen coating ELISA Agricultural Research Institute, New Delhi-11012, India
(A405nm= 1.2 to 1.8). The total RNA from the positive leaf samples was isolated and
tested through RT-PCR with the coat protein (CP) specific primers NS1 and NS2 which Email: [email protected]
the amplicon of 1065bp. The PCR product was further cloned in TA vector, confirmed
by restriction digestion, sequenced and submitted to NCBI data base (MG768974). Key words: Citrus tristeza virus; codon usage bias; citrus host; high frequency codons;
Sequence analysis of the CP gene showed 98 to 100% identity with Indian isolates codon usage adaptation
(KR260903, KR260902, and KP259608) followed by 82 and 92% similarity with Korean
isolates of SYMMV at nucleotide and amino acid levels, respectively. For further Abstract
confirmation mechanical sap inoculation was performed on French bean cv. Pusa Introduction: Citrus tristeza virus (CTV), a member of the aphid transmitted
Parvati with an extract from ELISA- positive soybean leaf sample which showed the Closterovirus, is the causal agent of the notorious Tristeza disease in several citrus
chlorotic blotches at 10-12 days post inoculation. These symptomatic leaves tested species worldwide. The codon usage patterns of viruses reflect the evolutionary
and confirmed for SYMMV with DAC ELISA and RT-PCR with CP primers. changes for optimization of their survival and adaptation in their fitness to the
Previously,SYMMV was reported on soybean from South Korea and North America external environment and the hosts. The codon usage adaptation of CTV to specific
More recently, SYMMV was reported on mungbean and urdbean from India. To our citrus hosts has not been studied, thus its role in CTV evolution is not comprehended
knowledge, this is the first report of natural infection of soybean by SYMMV in India clearly.
and suggests that this newly identified SYMMV may be adapting to other leguminous
host plant species. Methods: Therefore, to better explain the host-virus interaction and evolutionary
history of CTV, the codon usage patterns of the coat protein (CP) genes of 122 CTV
Development of gene deletion or point mutation based constructs of isolates originating from three economically important citrus hosts (55 isolate from
croton yellow vein mosaic virus to understand their suitability as replicon Citrus sinensis, 38 from Citrus reticulata and 29 from Citrus aurantifolia) were
vector studied using several codon usage indices and multivariate statistical methods.

Gurpreet Kaur, BikashMandal and Anirban Roy Results: The present study shows that CTV displays low codon usage bias (CUB) and
higher genomic stability. Neutrality plot and relative synonymous codon usage
Division of Plant Pathology, Indian Agricultural Research Institute, New analyses revealed that the overall influence of natural selection was more profound
Delhi – 110012 than that of mutation pressure in shaping the CUB of CTV. The contribution of high
frequency codon analysis and codon adaptation index value shows that CTV has host-
[email protected] specific codon usage patterns resulting in higher adaptability of CTV isolates
originating from C. reticulata (Cr-CTV), low in the isolates originating from C.
Begomovirus, Replicon, Cloning, Mutant, PCR aurantifolia (Ca-CTV) and C. sinensis (Cs-CTV).

It has been shown that genome of a plant virus can be modified in a way Conclusions: The combining codon usage analysis of CTV with citrus genealogy
so that it can efficiently replicate in the plant without producing symptoms suggests that CTV has evolved in C. reticulata, an evolutionary primitive Citrus species
or producing mild symptom. Hence, such modified virus genome based or other Citrus progenitors. The outcome of the study enhances the understanding
replicon can be used as a vector to carry a gene/gene fragment and deliver the factors involved in viral adaptation, evolution and fitness towards their hosts. This
that into plant. Such vectors are extremely useful to express any foreign information will definitely help to devise better management strategies of CTV.
protein in plant or to study functional behavior of any plant gene. Recently,
such plant virus based vectors werealso employed to deliver genome Development of single-tube multiplex polymerase chain reaction assay for
editing tools inside the plant. To utilize a plant virus as a replicon vector it simultaneous detection of C
is essential to know the minimal replicon size that allow their replication
and systemic movement inside the plant. With an aim to develop such [email protected]
plant virus based replicon vector, genome of a begomovirus(ssDNA),
Croton yellow vein mosaic virus(CYVMV) was modified and different gene Cotton leaf curl disease (CLCuD) caused by whitefly-transmitted monopartite (~2.7 kb
deleted and point mutant constructs were made from the infectious DNA-A) begomoviruses in association with beta- and alpha-satellites (~1.4 kb, both),
construct of CYVMV using PCR and cloning strategy. Besides that another is a serious constraint in entire cotton growing area of 1.1 Mha of Haryana, Punjab
construct containing the intergenic region, which has the origin of and Rajasthan in Northwest (NW) India. Betasatellite affects disease complex and
replication was made in pGreen II vector in tandem duplicates separated pathogenicity by suppression of host gene silencing, whereas, alphasatellite can
by a multiple cloning site with 6 restriction enzyme sites. All these reduce virus titre, preferentially betasatellite accumulation, and can attenuate
constructs were transformed first into DH5α cells, and then disease symptoms. Co-occurrence of the DNA-A and betasatellite molecules in CLCuD
electrotransformed in EHA105 strain of Agrobacterium tumifacience with infected cotton synergistically enhances disease symptoms. For detection of the virus
pSoup helper plasmid. To evaluate the constructs they wereagroinfiltrated and its associated satellite molecules, a quick, sensitive and cost effective detection
into Nicotianabenthamiana plants and the replication behavior of these method is required. With this objective a single-tube multiplex polymerase chain
constructs and symptom phenotype produced by themwere assessed. reaction (mPCR) assay was developed for simultaneous detection of CLCuD-
Such begomovirus based replicon vector will open up a new era for begomovirus (DNA-A), and its associated beta- and alpha-satellite molecules. Several
effective utilization of plant viruses as a gene delivery tool for plant. sets of primers for amplification of the virus and satellite molecules based on the
retrieved reference sequences from the GenBank were designed and tested
Synonymous codon usage pattern of Citrus tristeza virus reveals high codon considering various parameters, annealing temperature and template and primer
adaptability to Citrus reticulata, an evolutionary primitive citrus host concentration. Of them, three sets of primers targeting (i) complete coat protein (CP)
gene (750nt) of CLCuD begomovirus, (ii) complete βC1gene (363nt) of betasatellite
Supratik Palchoudhury, Shibu Das, Prosenjit Chakraborty and Kajal Kumar Biswas and (iii) partial Rep gene (445nt) of alphasatellite were standardized and used in
simplex (s) and multiplex (m) PCR. The constant amplified products were identified tolerant (H-88-78-1) and susceptible (Punjab Chhuhara) cultivars at different days-
on the basis of amplicon size in mPCR and were compared with sPCR. The amplicons post-infection. We showed that less abundance of viral replicative intermediate in
were cloned and results were confirmed by sequence analysis. The optimized mPCR tolerant cultivar may have a correlation with a relatively higher accumulation of virus-
protocol was validated by testing infected cotton samples exhibiting variable specific siRNAs. Further, a significant correlation was observed between siRNAs
symptoms collected from different areas of NW India. The mPCR will definitely save accumulation and altered methylation patterns in tolerant versus susceptible
time and reagent costs for diagnostics. This tool can be used for screening disease cultivars. These suggest that both viral DNA methylation and siRNA-mediated
resistance, quick diagnosis, virus indexing and epidemiological studies. degradation play an important role in conferring tolerance against ToLCNDV. Further,
we functionally characterized a (i) 26S proteasomal subunit RPT4a (SlRPT4) gene and
Characterization of Candidatus Phytoplasma australasiae-related strain associated (ii) U-box type E3 ligase (SlARM18) gene, which were differentially expressed after
with big bud and witches-broom disease of Tomato and Carrot in India based on ToLCNDV infection in tolerant cultivar. (i) The study showed that SlRPT4 protein binds
the studies of 16S rRNA and SecY gene to promoter region of ToLCNDV genome thereby hindering the expression of virus
genes which subsequently reduces viral replication and infection in tolerant cultivar.
V .Venkataravanappa*, P. Swarnalatha and M. Krishna Reddy Transient overexpression of SlRPT4 resulted in activation of programmed cell death
and antioxidant enzymes system. (ii) An increase in viral load upon silencing SlARM18
Dr.V.Venkataravanappa, Scientist (Plant Pathology), Division of Plant Pathology, in cv. H-88-78-1 was observed validating its role in tolerance. Upon virus infection,
Central Horticultural Experiment Station (CHES)-Chettalli-- 571248 WRKY41 was found to regulate the expression of SlARM18 by binding onto the W-
ICAR-Indian Institute of Horticultural Research (IIHR), Bangalore Karnataka, India box elements present within the promoter of SlARM18. Further, downstream,
SlARM18 interacts with viral AC4 protein and leads to its ubiquitinylation. Overall,
[email protected] present study highlights non-proteolytic function of SlRPT4 as well as SlARM18-
mediated ubiquitination of viral AC4 protein and their participation in defense-
Key words : Phytoplasma, RFLP, PCR, In-silco analysis pathway against virus infection in tomato.

Introduction: Tomato and carrot is one of the important vegetable crops grown
throughout the country under diverse agro climatic conditions. The crop is prone to Detection of six important begomoviruses and two different betasatellites in chilli
many fungal, bacterial, phytoplasmal and viral diseases. Among these, big bud and from Gujarat
witches-broom disease of tomato and carrot caused by phytoplasma is a looming
threat for tomato and carrot cultivation in India. [email protected]

Methods: Tomato and carrot plants showing big bud/witches broom were collected Zairah W, Pradeep Kumar, Anirban Roy, Bikash Mandal
from farmer field were identified based on the PCR using phytoplasmic-specific
primer pairs of 16S rRNA and SecY gene. The amplified PCR product was cloned and Chilli leaf curl has emerged as a serious viral disease in the cultivation of chilli in the
sequenced. different parts of India. In this study, nineteen chilli samples (ChGj1-19) showing leaf
curl and yellowing symptoms were collected from the farmer’s fields at Surat,
Results: The complete nucleotide (nt) sequence of 16S rRNA and SecY genes of Gujarat, India were analysed. In order to know the begomoviruses and their
tomato big bud and carrot witches broom phytoplasmas isolates showed maximum betasatellite associated with the leaf curl disease, PCR was conducted using
nt identity more than 95 %with Candidatus Phytoplasma australasiae (16SrII). Further degenerate as well as specific primers. The ten samples those showed positive with
In-silico RFLP analysis of 16SrRNA gene of tomato big bud samples showed similarity degenerate primers were further amplified using six begomovirus species specific
coefficient in the range of 0.68 to 0.95. Whereas carrot witches broom isolates primers [BM796F & BM797R for tomato leaf curl Bangalore virus (ToLCBaV); BM800F
showed similarity coefficient 0.93. Therefore the strains are significantly distinct from & BM801R for tomato leaf curl Gujarat virus (ToLCGuV); BM802F & BM803R for
other subgroups of Candidatus Phytoplasma australasiae. Further, the phylogenetic tomato leaf curl Joydebpur virus (ToLCJV); BM798F & BM799R for tomato leaf curl
analysis revealed that, they are closely clustered with Candidatus Phytoplasma New Delhi virus (ToLCNDV); BM794F & BM795R for tomato leaf curl Palampur virus
australasiae strains (16Sr II), specifically within the 16Sr II-D and 16Sr II-A subgroups. (ToLCPalV) and BM861F & BM862R for chilli leaf curl virus (ChiLCV)]. The PCR with the
species specific primers revealed prevalence of all the above begomoviruses and
Conclusions: The study highlights the identification of new strains of phytoplasmas importantly showed the association of multiple begomoviruses up to five different
associated with tomato and carrot in India. The association phytoplasma is one of the begomoviruses in a single plant sample. All the nine positive samples analysed for the
most important alarm signal for its production and serious threat cultivation these detection of betasatellite using universal beta primers (CLB36F & CLB37R). The five
crops. samples (ChGj14, 15, 16, 17 and 19) that showed the positive amplification with

Role of proteasomal pathway gene(s) in combating virus infection in plants Molecular screening and characterization of weeds as reservoirs of Begomoviruses
around leguminous crops
Manoj Prasad*
Sudeep Pandey1, Nagaraju, N2 and Girish TR3
National Institute of Plant Genome Research, Aruna Asaf Ali Marg, JNU Campus, New
1,2 Department of Plant Pathology, University of Agricultural Sciences, GKVK,
Delhi
Bengaluru-65
3 Sea6Energy Private Limited ,C-CAMP,NCB-GKVK Campus , Bengaluru-65
[email protected]

Tomato leaf curl disease (ToLCD), caused by strains of Tomato leaf curl virus (ToLCV), [email protected]
is a major constraint to tomato production. To understand the molecular mechanism
of virus tolerance, we studied the abundance of viral genomic replicative Key words: Begomoviruses, Weed hosts, Leguminous crops, RCA- PCR
intermediate molecules and virus-derived siRNAs generated by host plant in naturally
Abstract:
 Yellow mosaic disease (YMD) caused by Begomoviruses is a critical
constraint in the production of leguminous crops. A season-long survey was carried Introduction: Ubiquitination is a post-translational modification involved in various
out for YMD in six different leguminous crops and in associated natural weeds, across developmental processes and stress responses in plants. SlARM18 (U-box type E3
different regions of southern Karnataka during 2017 and 2018. As high as about 94.0 ligase) was found upregulated (~5 fold) during ToLCNDV infection in tolerant tomato
per cent of YMD incidence was recorded across six leguminous cropping systems cultivar. Given this SlARM18 was characterized for its role in tolerance against
surveyed. Among the 96 weed samples surveyed 54% were symptomatic while 45.8% ToLCNDV.
were asymptomatic. Both symptomatic and asymptomatic weed samples were PCR
screened using Deng primer, Mungbean yellow mosaic virus and Horsegram yellow Material: Silencing of SlARM18 was performed through VIGS. For promoter analysis
mosaic virus Coat protein gene specific primers. Among the asymptomatic weed deletion promoter constructs were generated in fusion with GUS-GFP reporter gene.
samples 55.20 per cent amplified through direct PCR, while in 18.75 percent samples EMSA was performed to validate DNA-protein interaction. Yeast-two hybridization
the PCR amplification was observed only after the enrichment of the template and BiFC were performed to identify interacting proteins. Ubiquitinylation assay was
through Rolling circle amplification. Out of the nine weed species commonly performed to validate ligase activity of SlARM18.
observed the weeds, Agertum conyzoides, Alternanthera sessilis, Commelina
benghalensis and Euphorbia geniculata were found abundantly. Further, sequence Results: Increase in viral load upon silencing SlARM18 in cv. H-88-78-1 was observed
analysis of PCR products amplified from weed samples revealed a close clustering validating its role in tolerance. Histochemical and fluorometric GUS assay revealed
with the Horsegram yellow mosaic virus isolateLima bean, Legume yellow mosaic involvement of two W-box elements within promoter of SlARM18 in tolerance
virus and Mungbean yellow mosaic virus isolate Nammakal, Moreover, a cross response. In-vitro DNA-protein interaction by gel shift assay indicates strong
infectivity test on blackgram and soybean crops using viruliferous whitefly, Bemisia interaction between WRKY41 and W-box elements present in the SlARM18 promoter.
tabaci that were fed respectively on weeds A. conyzoides and E. geniculate revealed SlARM18 was found to interact with AC4, viral protein and deletion of U-box domain
clear YMD symptoms. Overall, these data suggests a potential role of weed species abolished this interaction. In-vitro and in vivo Ubiquitinylation assay indicated that
on the epidemiology of the Begomoviruses in legume crop systems and thus useful SlARM18 comprises a strong ligase activity and it ubiquitinates viral AC4 protein.
towards developing future virus management strategies. Further AC4 was found to be the substrate for ubiquitinylation by SlARM18 leading
to its degradation via proteasome system.

Development of binary vector constructs with genes of Indian Citrus Ringspot Virus Conclusion: The present study provides novel clues on role of SlARM18 in conferring
toward identifica tolerance against ToLCNDV. SlARM18 ubiquitinates viral AC4 and it could be adopted
as a strategy for developing viral resistant plant by overexpressing SlARM18.
[email protected]
Funding: NIPGR and SERB.
Kinnow orange is one of the widely accepted fruit due to its vitamin C rich high juice
content. It is cultivated mainly in Punjab areas of India and has high export values. In Association of a Nucleorhabdovirus with vein clearing disease of cardamom in India
the recent past, Indian citrus ringspot virus (ICRSV) is reported to be widely-
distributed and known to infect Kinnow affecting qualitative and quantitative yield Bhat A I, Biju C N, Pamitha N.S
losses up to 100%. The infected trees produce the typical symptoms such as rings,
vein-clearing, vein-banding flecking and chlorotic pattern on the mature leaves. ICRSV ICAR-Indian Institute of Spices Research, Kozhikode 673012, Kerala, India
is classified into genus Mandarivirus and family Flexiviridae. It contains a single-
stranded + sense RNA genome size of 7.5 kb long which consist six open reading
frames (ORFs). Identification of ICRSV suppressor protein(s) which is able to interrupt email: [email protected]
the host defense by RNAi mechanism is required to understand the virus-host Key words: Cardamom vein clearing virus, transmission, small RNA sequencing, RT-
interaction. In this study, we collected the infected and healthy kinnow leaves based PCR
on typical symptoms from the ICRSV Punjab isolates maintained in glass house of IARI,
New Delhi. Total RNA from leaves were isolated using TRIzol reagent. Two-step RT- Abstract
PCR was performed with the gene specific primers containing the restriction sites The vein clearing (kokke kandu) disease caused by an unknown virus is an
targeting to clone into pCAMBIA binary 1302 vector. PCR amplified ORF2 of 678bp, important production constraint of the cardamom crop in India. The disease is
ORF3 of 330bp, ORF4 of 183bp and ORF6 of 669bp of ICRSV was cloned in binary characterized by chlorosis of the veins followed by rosetting, loosening of leaf sheath
vector pCAMBIA 1302. Positive clones were selected and DNA sequenced. The DNA and shredding of leaves; newly emerging leaves get entangled in the older leaves and
sequences were analyzed to be 99% identical with the previous isolates of ICRSV. The form hook-like tiller and hence the disease is locally known by the name kokke kandu.
positive binary vectors will be mobilized into Agrobacterium tumefaciens strain The infected plants decline rapidly with yield reduction up to 62-84% in the first year
GV3101 and agroinfiltration study will be performed in GFP expressing Nicotiana of peak crop and perish within 1-2 years of infection. In the present study causal virus
benthamiana 16c line to identify the suppressor protein. associated with the disease was successfully transmitted through aphid, Pentalonia
caladii. Small RNA (sRNA) sequencing of the aphid transmitted cardamom plant
WRKY41 mediated transcriptional regulation of a U-box type E3 ligase SlARM18 showed several contigs aligning with nucleorhabdoviruses. Two regions of the causal
leads to targeting of AC4 protein of Tomato leaf curl New Delhi virus virus covering 9806 nucleotides was amplified through RT-PCR using primers based
on the sequence of the contigs that mapped to the nucleorhabdoviruses. Cloning and
Arunava Mandal, Namisha Sharma, Priya Dulani, Shambhavi Sharma & Manoj Prasad* sequencing of the amplified product showed identities ranging from 30-62% in the
nucleocapsid (N), phosphoprotein (P), putative movement protein (P3), matrix
National Institute of Plant Genome Research, Aruna Asaf Ali Marg, JNU Campus, New protein (M), glycoprotein (G) and polymerase (L) genes of different
Delhi nucleorhabdoviruses thus confirming the identity of the virus as a distinct species
under the genus, Nucleorhabdovirus for which the name Cardamom vein clearing
[email protected] virus is proposed.

Key words: Geminivrus, Ubiiquitin-proteasomal system, Ubiquitin ligase, Tomato

Characterization, in silico prediction of begomovirus infected papaya (Carica Effects of biotic stress on the essential oil yield and quality of Mentha spp. and
Papaya) encoded microR screening diseas

[email protected] [email protected]

Leaf curl disease of papaya is caused by several species of Begomovirus, family Background Mentha (family Lamiaceae) consists of different species/varieties based
Geminiviridae. The leaf curl disease of papaya is transmitted by whitefly (Bemisia on flavor, and among them, three species of mints (Mentha arvensis, M. piperita, and
tabaci), and is single stranded monopartite in nature; however bipartite genome is M. spicata) are commercially cultivated in India for the essential oils. It is widely used
also reported. The disease is characterized by downward curling and cupping of in various pharmaceutical and confectionary industries. Since last decade apart from
leaves followed by vein thickening. Every year, India witness tremendous loss to its abiotic stress (water, salinity, light), biotic stress caused negatively influence the
papaya growing areas. Infected leaves were collected from papaya plants from Delhi survival, biomass production, oil yield, and quality. Comparative studies are reported
and its suburbs during 2016 and 2017 and virus DNA were cloned and characterized. here. Materials and Methods Different species of Mentha were studied in the
Agroinfectious clones were also developed. Bioinformatics study throws light on the experimental field of CSIR-CIMAP, Lucknow with reference to their health,
similarity and dissimilarity between the different isolates of papaya leaf curl virus that productivity, quality product and were also screened for the selection of improved
has infected papaya plant in various regions of India and sub continents. MicroRNAs genotype tolerant to fungus and virus infection. Conventional and molecular
(miRNAs) belong to class of endogenous small RNAs which suppress expression of approaches were applied to develop genotype with required characters. Essential oil
genes following cleavage or translational inhibition of target messenger RNAs. They extraction was carried out using Clevenger apparatus and further subjected for gas
are involved in a number of plant processes such as development, biotic and abiotic chromatography analysis to compare the yield and quality of oil in the healthy and
stresses. In this study, in silico approach is employed, high scoring miRNA-target pairs infected Mentha spp. Results Leaf curling, as well as necrotic spots and patches, were
satisfying rules of minimum free energy and maximum complementarities were found on the leaves of infected M. arvensis, M. spicata, and M. piperita. PCR results
selected. This study shows predicted papaya miRNAs which have the potential to showed the presence of viral and fungal infection. The main oil constituents were
target different genes, including AC4 region, which involve in gene silencing identified as limonene, menthone, menthofuran, menthol, carvone and methyl
suppression. Further next generation sequencing data reveals several new miRNA acetate. The chemical profile of M. arvensis oil showed an increase in menthol
from infected papaya leaf sample. content and decrease in limonene percentage in infected plant in comparison to
healthy while M. piperita showed increase in menthol content accompanied by an
Solanum whitefly Aleurothrixus trachoides (Back) can transmit Geminivirus ? increase in menthofuran. Some accessions/lines are identified as tolerant to
fungal/viral infection for the detailed studies. Conclusion Phyto-pathogens (viruses
K. Chandrashekar*, Ashutosh Rao, Savarni Tripathi and Rajverma and fungi) are responsible for the stress, cause changes in the quality and quantity of
the essential oil. Therefore to combat the effect of infection caused by fungi and
ICAR-IARI, Regional Station, Pune viruses, vigor genotype via breeding program is needed.

email id: [email protected] Molecular characterisation of Begamovirus associated with a new viral disease on
Clerodendrum serrat
Key words. Aleurotrachelus trachoides, Geminivirus, vector, Duranta sp
[email protected]
Background: Solanum whitefly Aleurotrachelus trachoides (Back). (Hemiptera:
Aleyrodidae) is reported to occur on tomato, capsicum, eggplant, tobacco, Duranta BACKGROUND Clerodendrum serratum (L.) Moon. (Verbenaceae) is an important
sp and several other weeds. The pest is native to the neotropics and presently medicinal plant growing in the tropical and warm temperate regions like Africa,
reported to occurs in several countries. In India it was first reported by Dubey and Southern Asia; Malaysia and distributed throughout in the forests of India and Sri
Sundararaj during 2015 from Karnataka and they described it as Aleurothrixus Lanka. It is traditionally valued and reported for treating pain, inflammation,
trachoides (Back). The damage caused by A. trachoides is mainly due to feeding on rheumatism, respiratory disorders, fever and malaria in India with a long history.
plants sap. In the year 2017 European and Mediterranean Plant Protection MATERIALS AND METHODS During routine field survey at CSIR-CIMAP in April 2018,
Organization declared A. trachoides as a non vector. However, infected plants show Clerodendrum serratum have been found to be suffering from green mosaic and
typical virus infection symptoms hence, the present study was undertaken to identify blistering on leaves as compared to healthy plants. To ascertain the presence of the
the possible occurrence of geminiviruses. begomovirus, total DNA was isolated from twelve symptomatic and one
Methodology: Samples of A. trachoides and its infested host plants Duranta sp. asymptomatic leaf samples of C. serratum by the CTAB method and subjected to PCR
tomato, capsicum, eggplant, Solanum nigrum and Ipomoea sp. were collected from using begomovirus CP specific primers (GemF/R). Further, the full-length genome was
Pune, Maharashtra. DNA was extracted from plant samples and from individual amplified by the RCA method and digested by different restriction enzymes. Obtained
whiteflies. The isolated DNA of whitefly and plant were tested for occurrence of bands were cloned and sequenced and sequences were analyzed by bioinformatics
Geminivirus using degenerate primers. tools . RESULTS Based on the visual virus-like symptom, about 30% of plants have
Results: PCR analysis using degenerate primers for geminiviruses showed been found to be symptomatic. Virus-like symptoms and presence of the whitefly
approximately 520 base pair amplicons as expected in 12 out of 20 Duranta sp population, a Begomovirus infection was suspected. PCR result showed ∼771bp
samples and 6 out of 40 whitefly samples collected from Duranta sp. None of the amplification in all symptomatic leaves (twelve) samples. PCR result supports the
other samples showed any amplification. The results indicated the presence of presence of the Begomovirus infection. Digested RCA product produced ∼2.7-kb
Geminivirus in Duranta sp and whitefly by degenerate primers and RCA amplification. band. Analysis of the sequence data showed 99% identities, and close phylogenetic
Further studies to characterise geminivirus transmission ability of A. trachoides is relationships with the DNA-A genome of Tomato Chilli leaf curl India
under progress virus(ToChiLCuIV) (FM877858) reported from India. CONCLUSIONS Based on high
Conclusion: The results suggest the presence of geminivirus in whitefly A. trachoides sequence identities and close relationships, the begomovirus isolated from C.
as well as in its host plant Duranta sp. Hence, there is a need for detailed studies to serratum was identified as an isolate of (ToChiLCuIV) infecting C. serratum from
understand and determine vector status of A. Trachoides. Lucknow, India. In the literature, there is no disease has been reported on C. serratum
so far. We, here, have reported Tomato Chilli leaf curl India virus infecting C. serratum
for the first time in India.
 nucleotides with potential ORF coding capacity. Transient expression of βV1 induced
Molecular characterization, pathogenesis and recombination studies of rapidly potential hypersensitive response and peroxides accumulation followed by cell death
evolving Begomoviruses in N. benthamiana plants. The βC1 transcript mapping revealed the ORF with a 5′ UTR
of 615 bp and 3′ polyadenylation signal 33 bp downstream to the ORF.
[email protected] Conclusion: Betasatellite transcript mapping identified multiple polyadenylated
transcripts. βC1 and putative βV1 ORF genes mapped and putative βV1 ORF induces
BACKGROUND Andrographis paniculata (Kalmegh) is a medicinal herb, well known for HR-mediated cell death.
its antioxidant, antidiabetic, anticancerous, anti-inflammatory, antimicrobial and
hepatoprotective properties. Begomovirus is the largest genus of the family Analysis of susceptibility to RNMV infected plants in response with M. oryzae and
Geminiviridae, infecting a large range of plants. Recombination is one of the major X.Oryzae, Xanthomonas oryzae pv. Oryzae and CMV
factors for the emergence of new species of begomoviruses and the expansion of its
host range. Although few fungal infections are reported, no viral infection had been Wagh, S. G. Shamim, A. Daspute, A., Bhor, S. A., Kobayashi, K., and Nishiguchi, M.
reported earlier on it. Our study is the first report of viral infection on A. paniculata.
MATERIALS AND METHODS During the survey of the Kalmegh growing areas, typical MGM, IBT, Aurangabad, INDIA.
symptoms made the preliminary detection of the viral infection. PCR performed using
begomovirus coat protein specific primers for the molecular detection of the virus. [email protected]
Full-length genome, amplified using PCR and RCA techniques, was sequenced and
characterized. Agrobacterium-mediated transformation was used to check the Key words: RNMV, VIRUS-FUNGUS INTERACTION
molecular infectivity of the virus in different hosts. Recombination studies were
analyzed using bioinformatics tools. ABSTRACT
RESULTS Typical virus-like symptoms such as leaf curling, vein clearing, yellowing Background: Rrice necrosis mosaic virus (RNMV) causing mosaic symptoms
were noticed on the infected plants of A. paniculata. Full length genome characterized by yellow flecks and streaks on lower leaves of rice plants was first
corresponding to 2.7 kb was obtained by PCR and RCA methods. Another genomic reported in Japan and then in India (Fuji, 1877, Ghosh, 1980, Badge et al. 1997). This
component of 1.3 kb corresponding to betasatellite was also detected. The complete virus has been classified as a member of the Bymovirus genus of the Potyviridae.
genome was sequenced and submitted in GenBank database. Based on the RNMV is transmitted by a fungus, Polymixa graminis.
nucleotide sequences, they were identified as Eclipta yellow vein virus (Acc no. Methodology: RNMV inoculated by growing rice seedling in soil infested with the
KC476655), Papaya leaf crumple virus (Acc no. KM359408) and a new species of fungal vector Polymixa graminis carrying RNMV. Wild type rice (Oryza sativa L. cv.
Begomovirus named Andrographis yellow vein leaf curl virus (Acc no. KM359406). Nipponbare) and the transgenic lines were used in this study. Oryza sativa cv.
Molecular infectivity assay revealed the presence of betasatellite, and it helped in Nipponbare is susceptible to both X. oryzae pv. oryzae and M. oryzae. The seeds of
symptom enhancement. Recombination events detected in all these three virus Asominori and Sensyou, which are resistant to X. oryzae pv. oryzae and M. Oryzae.
isolates. Results: We examine that RNMV virus infected plant were again inoculated
CONCLUSION The cultivation of A. paniculata is not sufficient to meet the demand of individually with pathogens Magnaporthe oryzae, Xanthomonas oryzae pv.
pharmaceutical industries. Heavy loss of such medicinally important crops by Oryzae,Cucumber mosaic virus. After inoculation lesion size analysed in both
begomoviruses should be managed. Magnaporthe Oryzae and Xanthomonas oryzae pv. Oryzae. Among them
Magnaporthe oryzae shown higher infection level, where CMV and Xanthomonas
Mapping and characterization of begomovirus betasatellite transcripts oryzae pv. Oryzae were moderate.The similar results confirmed by analyzing bacterial
biomass content and fungal DNA content, in case of CMV purified virus inoculated,
Reddy KK1, Chakraborty, S1 RNA content analyzed by q-RT-PCR
Conclusion: RNMV virus has moderate effect on yield of rice, but it may have role and
1
Molecular Virology Laboratory, School of Life Sciences, Jawaharlal Nehru University, in resistance breakdown and facilitation to divesting pathogens such as Magnaporthe
New Delhi-110067, India. Oryzae shown in this study. These findings indicate that RNMV infection alters plant
immunity which facilitates the other fungus and virus infection.
[email protected]
Animal Virology
Keywords: Begomovirus, betasatellite, βC1
Introduction/Background: Begomovirus associated betasatellites encodes for single Oral Presentation
multi-functional protein βC1, which plays a key role in betasatellite-mediated
pathogenesis. Mutagenic studies on betasatellite speculated the origin of different Vaccines and Vaccination Concepts in Bluetongue
transcripts other than predominant βC1 transcripts. The current study attempted to
identify and characterize the putative transcripts originating from Radish leaf curl A.B. Pandey
betasatellite (RaLCB-Rβ) upon co-inoculation with Tomato leaf curl New Delhi virus
(ToLCNDV-NA). Division of Biological Standardization, Indian Veterinary Research Institute, Izatnagar
Material and methods: Transcript mapping studies were performed by northern
blotting, primer walking and random amplification of cDNA ends (RACE). Bluetongue (BT) is a culicoides transmitted economically important disease of
Results: RNA gel blots of RNA extracted from NA+Rβ infected plants identified an domestic and wild ruminants. In India, disease was first reported in 1964, since then
uncharacterized transcript from the satellite conserved region (SCR) and the several outbreaks have occurred and it is now endemic in this country. In India, sheep
expression of the transcript enhanced as the disease progressed. Primer walking is the main species affected with the disease in the frequent outbreaks causing huge
identified multiple transcripts of various lengths ranging from 94 to 913 bp across the mortality. Other animals like goat, cattle, buffalo, antelope, dear, sambhar, camels
SCR and βC1 ORF. Since geminiviruses are reported to have bidirectional promoters, and llamas also get affected by/infected with the virus. Disease can take course of
RACE experiments were carried out for virion and complementary sense strands to asymptomatic to lethal forms depending upon the BTV serotype, species, breed and
further identify the nucleotide coordinates of transcription start and end points. 5′ age of animal involved. Disease causative agent is BT virus (BTV), belonging to the
and 3′ RACE uncovered a transcript on virion strand (βV1) between 234-584 Orbivirus genus of Reoviridae family. In sheep, the disease varies from acute to
chronic forms and sometimes there is subclinical infection. Affected sheep have reported. Swine being an amplifier host plays an important role in epidemiology of JE
fever, depression, anorexia, tachypnea, hyperemia of the lips and nostrils, salivation, and considered as a significant risk factor for transmission of virus to human beings.
nasal discharge, edema of the face and tongue, conjunctivitis, ulcers on oral mucosa, Seo-conversion in swine occurs 2-3 week before infection comes to human beings
rarely, cyanosis of the tongue, coronitis, laminitis, muscle weakness, lameness, and hence swine act as a suitable sentinel for prediction of JE outbreaks in human
consequently, animal stand with an arched back, dermatitis, torticollis and wool population. Recently, researchers have found vector free transmission of JEV in
breaks. Gravity of the disease problem can be understood from the fact that 22, out experimental pigs through oro-nasal route though it is yet to be proven in natural
of 27 serotypes known world-wide, are circulating in ruminants in India, which has field conditions.
varied climatic conditions across the length and breadth of the country having Though reported for the first time in 1955 in India, JE emerged as a major
competent vectors for disease transmission. Because of a large number of susceptible public health problem in India since 1973 when first major outbreak took place in
hosts, BTV serotypes and Culicoides vector, control of BT is not an easy task. The West Bengal and later the deadly outbreaks in year 1978 and 2005 in Gorakhpur
control strategy of bluetongue can be successful mainly through vaccination of region of Uttar Pradesh. Earlier maximum cases were reported from Uttar Pradesh,
animals with vaccine using right serotype of the virus. Different regions of the Assam, West Bengal and Bihar. Presently, JE virus has spread its realm to various
country, depending upon the number of virus serotype(s) prevalent in particular other states of India where earlier it was not reported. Several factors including
region will require the vaccine prepared from those respective serotype(s). An ideal climate change may be responsible for its spread. Government of India has included
BT vaccine is efficacious, safe, affordable, protective against multiple serotypes and vaccination against JE as part of the routine immunization under the Universal
enables the differentiation of infected from vaccinated animals. It is difficult to Immunization Program in 206 JE-endemic districts in the country. However, still there
prepare an ideal vaccine, however, a judicious balance will have to be made between is ample scope to increase vaccination coverage in the country.
efficacy and duration of immunity, safety for the animal and the environment, costs, Animal husbandry sector is actively involved in surveillance of JE in swine
applicability for multiple serotypes, and possibility for DIVA. Both the Live attenuated population particularly in endemic states like Uttar Pradesh. Screening of more than
vaccines and inactivated vaccines have been successfully used in China, South Africa, 3000 swine serum samples by ICAR-Indian Veterinary Research Institute, Izatnagar
Europe and other countries. A genetically engineered virus-like particle (VLPs) has from across the country revealed a sero-positivity of 32 per cent. IVRI has also
also been projected as a next generation vaccine. However, prevalence of multiple developed indigenous diagnostics for the detection of JE infection in swine.
serotypes within a limited geographical area and incidence of genetic and phenotypic Indigenously developed diagnostics and vaccines for human JE are also available in
drift during natural infection in vectors and hosts, generating neutralization-resistant our country. However, existing infrastructure facilities i.e. diagnostic laboratories for
phenotypic variants within a serotype make the circumstances further difficult. In the routine surveillance in human and veterinary field need to be scaled up. The
India, attempts have been made to develop inactivated BTV vaccines using BEI and sentinel surveillance must start throughout the country so that outbreaks in humans
Hydroxylamine. BEI inactivated BTV vaccine elicited both neutralizing and group are timely predicted and prevented. In addition, expansion of vaccination program,
specific antibodies whereas hydroxylamine inactivated vaccine elicited group-specific hospital upgradation and sanitation in rural and urban areas along with sentinel swine
antibodies. Both the inactivated vaccines showed reduction in clinical signs and surveillance are key elements in control and prevention of JE. Human vaccination
duration of viremia. An inactivated pentavalent vaccine (BTV-1, 2, 10, 16 and 23) has against JE has dramatically decreased incidence of JE and hence sustained efforts are
been developed and commercialized in the country. Modified live vaccines (MLVs) required to strengthen and expand JE vaccine coverage. Inter-sectoral collaboration
produce a viremia in animals which lead to further spread of the viruses and between human health, animal husbandry and other ministries to have a common
reassortment of their genome. Sheep rearing by the nomadic communities which are platform for surveillance and control program ensures better preparedness. Because
always on move will further restrict the use of MLVs. Many new vaccine candidates, of climate change, changing vector competence, shift in genotype of circulating virus,
ranging from subunits to replicating next-generation reverse genetics based vaccines, a complete relook into the epidemiology of JE is the need of the hour. The time has
have been developed with their merits and demerits and therefore the latest ones come to adopt One Health approach to control this disease in our country.
are yet to come to market. Hence, in Indian scenario, use of multivalent inactivated
vaccines is a good option for control of disease. Regular monitoring of the serotypes Current status of Peste des petits ruminants vaccination programme in India
of the virus circulating in the different parts of the country, their isolation and
identification to ascertain the serotype and to include it/them in V. Balamurugan*, G. Govindaraj, K.P.Suresh and Parimal Roy
monovalent/multivalent vaccine formulation will be proper strategy for successful
control of the disease. Various approaches to develop BT vaccines and issues of Indian Council of Agricultural Research-National Institute of Veterinary Epidemiology
vaccination will be discussed. and Disease Informatics (ICAR-NIVEDI), Post Box No. 6450, Yelahanka, Bengaluru -
560064, Karnataka, India

[email protected]; [email protected]
Japanese encephalitis: A zoonotic threat and our preparedness
Abstract
1 2
Ashok Kumar , Himani Dhanze and Chethan Kumar 3 Peste des petits ruminants (PPR), otherwise known as “goat plague” or
“Small ruminant plague” is enzootic in India as number of outbreaks have occurred
1IndianCouncil of Agricultural Research, New Delhi, India; 2ICAR- Indian Veterinary in the past and now being occurring regularly, round the year and most frequently
Research Institute, India; 3ICAR-Central Coastal Agricultural Research Institute, Goa, during the lean period throughout the country. In some Indian states viz. Karnataka,
India Andhra Pradesh, and Chhattisgarh the PPR outbreaks in sheep and goats have
declined after implementing the strategic mass vaccination control programme. The
Japanese encephalitis (JE) is a mosquito-borne zoonotic flaviviral disease decreased number of outbreaks in the recent past as well as changes in the disease
responsible for causing hundreds of fatalities in children every year in India. The virus severity patterns and distribution might be due to the existence of institutional
is maintained in complex cycle in nature wherein egrets and herons act as reservoir infrastructure, effectiveness of vaccine, timely vaccination and most importantly
hosts, mosquitoes of genus Culex as vector, swine as amplifier host and human beings effective planning and implementation of the vaccination programme. In overall,
as dead end host. Based on nucleotide sequences of envelope gene, JEV has five fixed strategies may not work for all the states or region or the countries. However,
genotypes (I–V) and only one serotype. All known JEV strains isolated in India until in the mass vaccination in pulse polio model with two to three cycles of vaccination,
2007 belonged to genotype III. Later, the presence of genotype I in 2009 in Gorakhpur with each cycle of covering entire population of sheep and goats initially,
region (Uttar Pradesh, India) and subsequently in 2010 in West Bengal has also been subsequently bi-annual vaccination covering the naïve young population in a
predesignated stipulated period for two years, followed by entire population, will The ecology of Assam represents a classical example of reasons for upsurge of JE in
have a tremendous impact on the control of PPR outbreaks in sheep and goats and human and animal population. JE occurred in sporadic outbreaks or epidemic forms
subsequent eradication. Vaccinating the migratory population at the check post or in upper Assam since 1976, but has become endemic and highest number of human
border regions of the states or inter-state border or in the place of entry or place of casesare reported from Assam since 2011. It may be attributed to frequent floods,
trade market of animal through transport from other states are to be targeted for increasing rice & pig farming, and abundance of mosquitoes. We reported 22.99% of
mass vaccination as and when required. Sharing the experiences on the PPR 335 pigs JEV sero-positive during 2015-16. There was significantly higher JEV positivity
vaccination strategies adopted by some of the successful states in India may provide in pigs from Sivasagar district (31.31%) than in Kamrup Rural district (11.51%). With
directions for other Indian states or other countries of similar socio-economic and the global warming, there has been increase in JE vector proliferation and longevity.
small ruminant rearing pattern to vaccinate and control PPR. The changing agricultural practices such as irrigation, increase in irrigated rice-
Keyword: PPR, Strategies, Vaccination, Control programme, India production area has resulted in increase in JE outbreaks. Climate change is also
influencing migration patterns of birds, which may result in introduction of JEV into
Epidemiology of Japanese Encephalitis in Animals: An Indian Perspective
new areas.
Baldev R. Gulati
In contrast to pigs, sporadic cases of JE occur in horses in India. Sero-surveillance
Principal Scientist, ICAR-National Research Centre on Equines, Hisar, Haryana among equines in 13 different states of India revealed maximum JE positivity in
Manipur followed by those in Gujarat, Madhya Pradesh and Uttar Pradesh. JEV has
Japanese encephalitis (JE) is an important and widespread mosquito-borne zoonotic
been isolated from a horse showing neurological signs of encephalitis from Haryana.
viral infection of horses, pigs and humans. JEV causes reproductive disorders in
Phylogenetic analysis of the full-length sequence indicated that the JEV strain isolated
pregnant sows and encephalitis in horses. JE is a disease of rural agricultural areas
from horse in India belongs to genotype III.
where vector mosquitoes proliferate in close association with pigs, wading birds, and
ducks. The principal vector species is Culex tritaeniorhynchus, which is present in There are still no specific drugs available to treat JEV infection. Prevention of JEV is
great density in rainy season in both tropical and temperate regions. This paper gives based on mosquito control, avoiding mosquito bites, pig and human immunization.
an overview of epidemiological factors associated with the spread of JE infection in Vector control is one of the most important aspects of the prevention of JE spread.
animals, current diagnostic techniques and options for controlling JE infection in Larvicides and insecticides help in controlling mosquitoes in paddy fields. Multiple
animals. types of JE vaccines are available for humans including mouse brain-derived
inactivated, cell culture-derived live-attenuated, cell culture-derived killed-
JEV is a plus-sense, single-stranded RNA of ~11 kb belonging to the genus Flavivirus
inactivated, and genetically engineered live-attenuated chimeric vaccines. Other
in the family Flaviviridae. Based on E gene phylogenetic analysis, JEV strains have
experimental vaccines, including recombinant poxvirus vaccine, DNA-based vaccine
been classified into five genotypes (G1-V). Although GIII is most widely distributed
are also under various stages of development for human use.
genotype in human population, GI is emerging predominantly in South Korea,
Thailand and China and India. In animals, GIII is widely distributed in pigs from several JE in livestock is a notifiable OIE-listed disease that could affect trade between
countries, including India while GI from China, Cambodia, Korea & Thailand. Equine countries. Vaccination is recommended for the control of disease in animals for
JEV isolates from Japan, Hong Kong and Taiwan have been typed as GI, GII and GIII. reducing abortions in pigs or protecting high value race horses. In South Korea, a live
We isolated JEV from a horse in India that belongs to GIII. attenuated JE vaccine (Anyang 300 strain, G3) has been used in swines for more than
30 years. This has proven highly effective at reducing incidence of disease in pigs. A
A definitive diagnosis of JE depends on isolation or demonstration of virus in sick or
formalin-inactivated vaccine derived from porcine kidney cells or chick embryo cells
dead animals. The routine laboratory diagnosis of JEV infection is based on culture
and live-attenuated vaccines derived from porcine kidney cells are used in swines or
and serology followed by identification by RT-PCR. Virus isolation is done by intra-
horses in several Asian countries. Vaccination of horses is effective in protecting
cerebral inoculation of clinical specimens in suckling mice or passaging in Vero,
against disease, particularly high value animals such as racehorses that travel to JE
BHK21, LLCMK2, C6/36. Molecular diagnostic methods for diagnosis include RT-PCR,
endemic regions. In India, no vaccination has been practiced for animals, including
fluorogenic (TaqMan) or SYBR green-based real-time PCR and loop-mediated
horses and swine. Since pigs are the amplifying host for the JEV, vaccination in pigs in
isothermal amplification (LAMP). Serological tests in animals include virus
the JEV-endemic areas will help in curtailing the source of JEV infection. Vaccinations
neutralization (VN), haemagglutination inhibition (HI), complement fixation (CF) and
of pigs will break the mosquito-pig-human transmission cycle and thus helping in
ELISA and IgM-capture ELISA.
control of JEV.
JEV is transmitted in a zoonotic cycle among mosquitoes and vertebrate-amplifying
Intensification and expansion of irrigated rice production systems over the past few
hosts, chiefly pigs and wading birds. Pigs serve as amplifying hosts because they
years have contributed in increased JE incidence. Effective JE control is possible by
develop viremia that remains high enough to infect mosquitoes for up to 4-14 days.
means of effective surveillance systems and timely vaccination of persons or animals
Humans and horses are considered as the dead-end hosts as they have brief periods
at risk. The animal diagnostic facilities for JE in India need to be strengthened for
of viremia with low virus titres. The ardeid wading birds (heron, egrets) are
timely diagnosis and forecasting of the impending outbreaks. Newer diagnostic
considered the primary enzootic hosts of JEV and play important role in epizootic viral
methods are needed for monitoring the spread of the disease in human, animal and
amplification. Mosquitoes that commonly transmit JEV are most heavily
vector population. There is no specific treatment for the disease. There is need for
concentrated in rural, rice growing areas and feed most actively in the late afternoon
development of safe and effective vaccines for use in animals. Strategies for the
and early evening. Although JEV has been isolated from over 30 species, paddy-
reduction of virus burden in ecology by controlling vector population and relocating
breeding mosquitoes of the Culex vishnui subgroup, particularly Cx.
pig populations away from human habitation may be considered for effective global
tritaeniorhynchus, are the major vectors of the virus. Till recently, JEV transmission
control of JE.
has been exclusively described as being mosquito-mediated. However, recently it has
been demonstrated that JEV can be transmitted between pigs in the absence of
arthropod vectors. Pigs shed virus in oronasal secretions and are highly susceptible Highly pathogenic avian influenza - an update on genetic diversity, geographical
to oronasal infection. Further, virus persists in pig tonsils for at least 25 days. These spread and its control and containment
findings could have a major impact on the ecology of JEV in temperate regions with
C. Tosh, S. Nagarajan, Manoj Kumar
short mosquito seasons. Persistence of viral infection under low environmental
temperature has been demonstrated in insectivorous bats. ICAR-National Institute of High Security Animal Diseases, Anand Nagar, Bhopal - 462
022, India
 last decade there is enormous progress in the development of molecular techniques
which have not only accelerated disease diagnosis, but also facilitated to differentiate
H5 and H7 Highly Pathogenic (HP) Avian Influenza (AI) viruses represent a threat to
virus strains and pinpoint the source of infections. Several PCR assays have been
animal health, human health and economy. Currently circulating H5 HPAI virus,
developed for rapid detection of the pathogens. More than one of these pathogens
emerged in 1996 in China, is evolving and continuing its spread by infecting poultry,
can be detected simultaneously by the use of multiplex PCR. Reverse transcriptase
other birds and occasional human infection. Following spread of H5 virus in other
loop mediated isothermal amplification (RT-LAMP) along with its modifications has
Asian countries, it had spread to Europe, Africa and North America, and wild birds are
been reported for detection of the viruses in less time with limited facility. Despite
considered as the cause of intercontinental spread. The virus has evolved into
the progress, newer diagnostic tests developed in the country need validation, inter
multiple genetic clades (namely 0-9), and recently it has undergone reassortment
laboratory comparison or external quality assurance and harmonisation. Besides,
with other low pathogenic AI (LPAI) viruses to generate novel H5N2, H5N3, H5N6 and
there is a need to work with one world,one health concept for managing the
H5N8 subtypes. The H5 virus is endemic in Bangladesh, China, Egypt, Indonesia and
economically important diseases of pigs and to develop user friendly, cost effective
Vietnam. In 2013, a novel H7N9 LPAI virus emerged in live poultry markets and is
and field based diagnostic for rapid and specific detection of the diseases in India. In
responsible for multiple epidemic waves of human infections in China. In 2017, the
the present paper advances in diagnosis more particularly the molecular tests
H7N9 LPAI virus mutated to HPAI virus in poultry and is spreading westward in China.
developed for the diseases of pigs in India along the management of the diseases will
Two major genetic lineages of H7N9 virus (Pearl River Delta and Yangtze River Delta)
be discussed.
has been identified. Besides H7N9, other H7 subtypes circulating in recent years are
H7N2 (Australia), H7N3 (Mexico) and H7N7 (UK). Majority of the countries are
Key words: Pigs, diagnosis, management, molecular tests, endemic and emerging
following detect and culling policy for control and containment of HPAI. However,
disease
vaccination is advised as one of the components of a comprehensive control plan.

India, since its first report in 2006, has reported 159 H5 HPAI outbreaks (H5N1-126 RNA Viruses: Greatest Global Threat and One Health Solutions
and H5N8- 33) in 22 States/ Union Territories till March, 2018. Genetic analysis of the
viruses indicated multiple genetic clades (H5N1: 2.2, 2.2.1, 2.3.2.1a and 2.3.2.1c, and Prasad Minakshi a*b, Basanti Brar1a, Upendra P Lambe1a, Koushlesh Ranjan4a , Gaya
H5N8: 2.3.4.4) of the viruses indicating multiple introductions of the viruses. Prasad4a, Harimohan5a, Jinu Manoj6a
Currently, India is following “detect and culling” policy for control of HPAI. However,
a,b, 1a
transboundary nature of the virus warrants international collaboration with Department of Animal Biotechnology, LLR University of Veterinary and Animal
neighboring countries for effective control and containment of HPAI. Sciences, Hisar-125 004, Haryana, India.
2aCIRB, Hisar; 3aDepartment of Veterinary Physiology, COVAS, KVASU, Pookode,

Wayanad- 673576, Kerala, India; 4aSVP University of Agriculture and Technology,

Advances in diagnosis and management of economically important endemic and
Meerut, India; 5aIPVS, LUVAS, Haryana, India; 6aRVDEC Mahendergarh, LUVAS,
emerging viral diseases of pigs
Haryana, India

Dr. Dilip Kumar Sarma

[email protected]

Ex.Director, ICAR-NRC on Pig and Professor

RNA viruses can be divided into various classes on the basis of their genome polarity:
positive-strand RNA viruses (PSVs) and negative-strand RNA viruses (NSVs), linear or
Department of Microbiology, Faculty of Veterinary Science, Assam Agricultural
segmented based on genome arrangements. RNA viruses reproduce less accurately.
University, Khanapara, Guwahati-781022
They usually are deficient in proofreading and more prone to mutation rates of any
other organism on planet earth. These mutation rates mean that a large complex
Email: [email protected]
genome is not possible because their high error rates would cause offspring requiring
a large gene set to be nonfunctional. RNA viruses, therefore, have small genomes and
Pig rearing is one of the fastest growing livestock sector worldwide as it is an
lesser genes. The benefit of such a high error rate is that RNA viruses are able of
important source of rural livelihood and valuable source of animal protein. Pigs are
rapidly outmaneuvering the host immune system. The approach of RNA viruses is
also facing myriads of problems particularly the viral diseases, which have the
usually fast reproduction and moving on to a new host. Because they have less
capacity to decimate entire herds. Pigs are also regarded as mixing vessel, which gives
complex relationships with their hosts, RNA viruses are much more capable of
opportunity to evolve new virus strains capable to cause pigs and human health
moving to new host species. The ability to move to new hosts reduces the selective
problems. Despite huge opportunity and potentiality of the piggery the growth of this
pressure to not harm the host, and many RNA viruses are more pathogenic. In the
sector is slow in India. A number of endemic and emerging viral diseases like classical
present globalized world with overflowing travels and trades, human health has been
swine fever (CSF), foot-and-mouth disease (FMD), porcine circovirus type 2 infection,
increasingly threatened due to incidences of emerging and re-emerging infectious
porcine parvovirus infection, swine pox and porcine reproductive and respiratory
viral diseases. The most important sources of novel human pathogens are mainly
syndrome (PRRS) etc. are causing constant threat to the pig population of India.
farm mammals, poultry, to some extent wild animals and arthropods. It is estimated
Besides, with the increasing trend of human population, more demand for pork,
that zoonosis constitute about 60% of the ‘known’, and up to 75% of ‘emerging’ new
globalisation, climate change and porous international borders particularly in the
human infections. Over the past several decades, sporadic and often isolated
North Eastern Region may lead to emergence of newer diseases for which newer
outbreaks of emerging human diseases have led to the discovery of a diverse array of
tools will be important to predict, detect and mitigate in future. In order to reduce
novel, highly pathogenic viruses not limited to Filoviridae, Arenaviridae,
the risk associated with the endemic and emerging viral diseases more robust disease
Annuloviridae, Picobirnaviridae, Bunyaviridae, Paramyxoviridae, Coronaviridae and
surveillance, rapid and sensitive diagnostic tests, quality vaccines and improved
Flaviviridae families. Emergences as well as re-emergence of several viral infections
management are essential along with strengthening of veterinary services and
namely Nipahvirus, Hantaviruses, Chikungunya, Human Enterovirus-71, Influenza,
infrastructure and educating the pig rearers about the diseases and importance of
Chandipura, Crimean Congo, SARS, Picobirnavirus, Coronavirus, Buffalopox, Dengue
better management.
and Japanese Encephalitis viruses has-been reported from developing countries
Conventional diagnostic tests are widely used, but most of the tests are time
including India. The transmission of agents depends upon long time interactions
consuming, less sensitive and laborious. It is also difficult to accurately and
between people and domestic animals or nearby wildlife. The key to understand the
confidently predict viral infection status based on the clinical symptoms. During the
emergence and re-emergence viruses is to know the ‘host-pathogen-environment’
relationship in virus evolution. It is worth to mention that increasing number of cases PI animals, movement controls for infected herds, strict biosecurity, surveillance and
reported globally is alarming, indicating humans as source of transmission of vaccination as an additional tool. Although vaccination has reduced the clinical
infectious viruses to domestic and wild animals. Recent examples include influenza A, outcomes and spread of BVD, over the years vaccination alone has not resulted in the
astrovirus, enterovirus, human adenovirus, human metapneumovirus, human elimination of BVDV related clinical disease or a significant reduction in BVDV losses.
corona, hepatitis A, norovirus, rotavirus etc. The most affected animals are wild Emergence of novel BVD viruses, such as mixed triple highly virulent BVDV-2c has
followed by livestock and companion animals. Recent isolation and genetic analysis been reported recently with high mortalities in cattle in Germany and Netherlands,
of human H1N1 and H3N2 influenza viruses from pigs in China, human rotavirus from where coexistence of 3 distinct genomic variants was evident with unusual mutations
sheep and buffalo from India and developed countries provide strong evidence of in two variants. Additionally, co-circulation of novel and highly divergent BVDV-3
reverse zoonoses where transmission of viral agents is not restricted to developing viruses in cattle has recently been reported from India and BVDV-3 associated
countries, indicating a worldwide disease threat. Various contributing factors namely mucosal disease cases in some countries are on the rise. Although BVDV spread from
rapid movements of humans and animals, overall increase in industrial animal wild ruminants to cattle has a low to moderate risk, risk is high for BVDV transmission
production, and conflict of habitats of humans and wild animals are adding to from sheep and goats. Emergence of other ruminant pestiviruses include Tunisian
opportunities for humans to cause reverse zoonoses. sheep pestivirus, Turkish sheep pestivirus and Pronghorn antelope pestivirus.
Zoonotic infectious diseases have been an important concern to Furthermore, emergence of border disease virus (BDV) in wild ruminants and
humankind for more than 10,000 years. Today, approximately 75% of newly emerging increasing reports of natural transmission of BDV from sheep and goats to cattle
infectious diseases (EIDs) is zoonoses that result from various ecologic, leading to generation of PI cattle and further cattle to cattle transmission pose
socioeconomic, genetic, anthropogenic and climatic factors. These interconnected additional challenges for control in BVD free countries. Virological screening for
driving forces make it complicated to envisage and to prevent zoonotic EIDs. While identification and elimination of PI animals has been the hallmark for BVD control.
considerable improvements in various areas of environmental and medical However, acutely infected animals with highly virulent BVDV strains have recently
surveillance, clinical analytical methods and medical practices have been achieved in been observed as a high risk of spread and should not be ignored. The present lecture
current years, EIDs remain a foremost global concern particularly in less developed will focus mainly on emergence of novel BVDV strains and other ruminant
regions. The current Ebola epidemic in West Africa is a tremendous harsh reminder pestiviruses, and its implications on BVD control at global level so that adequate
of the role animal reservoirs play in public health and reinforces the vital need for changes in monitoring and control strategies can be formulated and implemented.
globally operationalizing a ‘One Health approach’. The complex nature of zoonotic
diseases and the limited resources in developing countries are a reminder for EMERGENCE OF TRANSBOUNDARY VIRAL DISEASES OF PIGS IN NORTH EASTERN
implementation of Global One Health in low-resource settings is essential. The STATES: A THREAT FOR INDIA’S MAIN LAND
Veterinary Public Health and Biotechnology Global Consortium launched the
International Congress on Pathogens at the Human-Animal Interface (ICOPHAI) in N. N. Barman*, A Sen, T Dutta, D. Hemadri, S. S. Patil, Suresh, B. C. Bera
order to tackle significant challenges and desires for capacity building. In One Health
implementation, four key capacity-building step needs: establishment of sufficient * Department of Microbiology, College of Veterinary Science, Assam Agricultural
science-based risk management policies, skilled-personnel capacity building, University, Khanapara, Guwahati, Assam- 781022, India
accredited veterinary and public health diagnostic laboratories with a shared
database, and improved use of existing natural resources and implementation. The [email protected]
future scientific research efforts must be aimed in areas of zonotic and
zooanthroponosis, for better understanding and transmission of various viral agents
Key words : North eastern region, pigs, transboundary, viral disease.
amongst various hosts resulting in accurate diagnosis and control measures.
ABSTRACT
Multidisciplinary approaches such as ‘One Health One World’ is the need of the hour
The north eastern region of India shares highly porous and sensitive frontiers with
to mitigate these problems.
China, Myanmar, Bangladesh and Bhutan. Again, rapid globalization opens frequent
international movements of men/material/animals, including vector population and
Emergence of novel bovine viral diarrhoea viruses and other ruminant pestiviruses:
bears an ever increasing threat of incursions of exotic, trans-boundary diseases
Implications on BVD control
crossing the borders to India’s main land through this North East corridor. A
retrospective as well as prospective monitoring of emerging viral diseases of pigs is
Dr. Niranjan Mishra
being carried out under DBT sponsored project involving three core NE laboratories,
four National Labs and eight directorates of animal husbandry and veterinary in NER.
Principal Scientist, ICAR-National Institute of High Security Animal Diseases, Anand
Epidemiological data, various clinical samples and tissue samples of pigs were
Nagar, Bhopal, Madhya Pradesh, 462022, India
collected from eight states of NER. To generate disease incidence as well as sero-
prevalence data of various infectious diseases of pigs, meta-analysis was conducted
E-mail: [email protected]
using the R Open source scripting software 3.4.3. Various molecular tools were used
to confirm classical swine fever (CSF), porcine reproductive and respiratory syndrome
Bovine viral diarrhea virus (BVDV) is an important pathogen of cattle with a global
(PRRS), porcine circovirus-associated disease (PCVAD), swine influenza (SI), torque
distribution and causes major economic losses due to a variety of disease syndromes
tenosus virus (TTSuV) disease and swine pox diseases. Sequencing was done to
including the highly fatal mucosal disease. BVD is caused by BVDV-1, BVDV-2 and
understand molecular epidemiology of all emerging swine viruses. In appropriate
BVDV-3 with considerable genetic and antigenic heterogeneity of BVDV strains.
conditions, isolation of viruses was done. All sera samples were subjected for
BVDVs belong to the Pestivirus genus within the Flaviviridae family that also
prevalence of antibody in unvaccinated population using in-house optimized i-
comprises the genera Flavivirus, Hepacivirus and Pegivirus. Significant differences
ELISA/or standard commercial kits.
exist in the virulence of BVDV strains. BVDVs are highly successful to persist and
Classical swine fever is endemic in NER. However, new geno-groups are
spread in their host populations due to their unique ability to produce persistent
emerging from neighbouring countries. Out of reported occurrence of CSF, annual
infection through evasion of adaptive immune response and innate immune
outbreaks were recorded as 24.46. Sero-prevalence of CSFV specific antibody
response. Recent advances in diagnostic methods, nucleotide sequencing and
indicated 31.00% in meta analysis. Further, CSFV genogroups 1.1, 1.2, 2.1 and 2.2
computer-assisted phylogenetic analyses have so .far identified twenty-one BVDV-1
were circulating in NER. Emergence of porcine circovirus-associated disease (PCVAD)
subtypes (BVDV-1a to -1u), four BVDV-2 subtypes (BVDV-2a to -2d) and four BVDV-3
of pigs caused by Circovirus 2 was confirmed. Sofar, 124 incidences were found
subtypes (BVDV-3a to -3d). Successful BVD control is highly dependent on removal of
positive for Circovirus-2 and also authenticated by gene sequence analysis.
Serological survey showed that 43.00% pigs was positive for porcine circovirus. Again, Key words: Next generation sequencing, NGS, Reovirus
porcine reproductive and respiratory syndrome (PRRS) was identified as new
emerging pathogen of pig in the state Meghalaya followed by in Mizoram. The Expression of VP2 protein of bluetongue virus serotype-1 in baculovirus system and
incidence was recorded as 13.03% in these two states. Authenticated sero- its potential use as a serotype specific diagnostic antigen
prevalence of PRRS was 2.00%. All the samples were screened by RT-PCR assay for
detection of ORF7 and ORF4 genes and sequence analysis showed close grouping with CHAND K1, SHARMA P2, BISWAS S K3, LOHUMI A4 AND RAMAKRISHNAN M.A5.
PRRSV isolates of genotype 2 (North American type) from China. Swine influenza virus
was detected in pigs of NER. Out of 69 nasal swabs from pigs, 13 samples showed Division of Virology, ICAR-Indian Veterinary Research Institute, Mukteswar Campus,
positive amplification of matrix gene of influenza A virus. Sequencing of PCR Nainital -263 138, Uttarakhand, India
amplicons (244bp) revealed 90-99.9% identity to the influenza A viruses. In these
study, torque tenosus virus (TTSuV) a small non-enveloped virus, containing single- Email: [email protected]
stranded, negative sense circular DNA was identified in limited NE states. The virus
commonly acts as cofactor with PCV and PRRS. Out of 51 samples 4 and 8 samples Introduction: Bluetongue (BT) is an arthropod-borne disease of
were found positive for TTSuV1 and TTSuV2, respectively. Along with climate change domestic and wild ruminants, caused by bluetongue virus (BTV). At present,
swine pox has been re-emerged in North Eastern States. A total of 9 outbreaks were worldwide 27 distinct BTV serotypes have been recognized. The BTV genome
confirmed as swine pox. On sequence analysis, SWPV isolates shared maximum consists of ten segmented (dsRNA) which codes for seven structural and four
identity (99–100%) with the previously reported SWPV isolate from India (AVA121) non-structural proteins. BT is enzootic in India and amongst the 23 serotypes
as well as from other countries. Swine pox virus showed a distinction from other reported to be circulated in the country, BTV-1 is one of the major concerns.
poxviruses. Isolation of CSFV, PCVADV, SIV, SWPV was done successfully. Detection of serotypes is necessary for taking up effective control measures
Confirmation of these emerging viral pathogens of pigs circulation in NER is a valuable including vaccination. VP2 is the most variable of the BTV proteins and is the
information and warrants preparedness to curb spreading of these viruses to India’s major serotype determinant. Therefore, the experiment was designed to
mainland. produce recombinant VP2 protein and evaluate its diagnostic potential for
detection of type-specific antibody against BTV-1 serotype.
Role of Next Generation Sequencing in discovery of neglected viral pathogens of Method and Results: The truncated N-terminal BTV (1)-VP2 (aa636), gene of
family Reoviridae BTV serotype-1 was amplified, cloned and expressed in baculovirus expression
system. For the recombinant baculovirus expression in Sf21 cells, the VP2
Yadav P1, Shete A2* and Mourya D3 gene, cloned into an appropriate baculovirus expression vector (pFastBac™)
and transposed into the bacmid DNA of DH10Bac™. The recombinant bacmids
1
Scientist E and Head of department, Maximum Containment Facility,ICMR- NIV are then isolated and used to transfect Sf21 cells. The analysis of the expressed
Pune; 2Scientist D, Maximum Containment Facility ICMR- NIV Pune; 3Director, Indian recombinant BTV (1)-VP2 proteins on SDS polyacrylamide gel revealed the
Council of Medical Research, National Institute of Virology, Pune protein corresponding to the theoretical molecular weights of BTV(1)-
VP2(aa636). By separating the soluble and insoluble proteins of the Sf21 cell
Maximum containment facility, Microbial Containment Complex, ICMR- National lysate, also revealed that the recombinant BTV (1)-VP2 proteins were soluble
Institute of Virology, Pune 410021, India since they were only found in the supernatant fraction. Further, the diagnostic
potential of the recombinant VP2 was evaluated by indirect ELISA using HIS
[email protected], [email protected] against BTV-1 and serum sample of sheep and goat origin.
Conclusion: The recombinant VP2 antigen based i-ELISA will be a good choice
Introduction: Detection of novel pathogenic viruses in the clinical samples has special for detection of type specific antibody against BTV-1 in the sero-
significance in the public health owing to the fact that many recent disease outbreaks epidemiological studies. The type-specific i-ELISA may be a suitable alternative
have been reported from the novel pathogens. The Next-Generation Sequencing to the cumbersome process of conventional serum neutralization assay.
(NGS) has revolutionized the genomic approach for detection of the viruses. The Keywords: BTV-1, VP2 protein, baculovirus
unbiased approach of sequencing implied in NGS has a major role in the detection of
unknown or uncharacterized pathogens. Molecular epidemiology and genetic analysis of coronaviruses from large and small
ruminants
Methods: Total RNA extractions was performed using supernatants and pellets Sircar S1*, Malik Y. S2, Bhat S3, Agarwal R. K4, Abhisek5, Sankar M6 and Muthuchelvan
obtained from cell culture and mice brain suspensions. Libraries were prepared using D7
the TruSeq LT Stranded mRNA Library Preparation Kit. The denatured libraries were
loaded onto Illumina Miniseq mid-output 150 reagent cartridges. The resulting fastq 12345ICAR - Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh–
files analyzed using CLC Genomic Workbench. 243122, India.
67
ICAR - Indian Veterinary Research Institute, Mukteswar, Nainital, Uttarakhand–
Results and Conclusions: We have characterized four previously un identified 263138, India.
reoviruses from India. Wad Medani virus (WMV), isolated in 1954 from a Hyalomma
marginatum ticks pool from Pune, Kammavampattai virus (KVPTV) isolated in 1963, [email protected]
from Sturnia pagodarum, Brahmani Myna bird from Tamil Nadu. One tick isolate was
identified as Kundal virus a member of the coltivirus genus. Equine encephalosis virus Keywords: Coronavirus, bovine, small ruminants, epidemiology, sequence analysis
(EEV), a reovirus, isolated from a dead horse in 2008 , first time from India. Introduction
Identifications of previously uncharacterized isolate lead to design assays for these Coronavirus (CoV) is a significant cause of diarrhea in calves and goat kids, winter
viruses. Molecular and serological assays developed can be used for further studies. dysentery in adult cattle, and also respiratory infections in calves. Despite its global
Diagnosis of the pathogenic viruses using NGS technologies can play a vital task in connotation, epidemiological data pertaining to CoV infection in animals from India
dealing with the global viral epidemics. This in turns can help in preparing for the is occasional. Therefore, a survey was conducted to detect CoV infection in dairy
emerging viral infections which can emerge as a major public health challenge. calves and goat kids and analyze the virus isolates genetically.
Material and Methods Keywords
From April 2017 to March 2018, 151 fecal samples from cattle (n=65), buffalo (n=22) CRISPR/cas9, Herc5, PPRV, gRNA
and caprine (n=64) were screened for CoV infection, respectively using RT-PCR based
detection system targeting the most consensus region from nucleocapsid gene (N)
segment for the CoV. Amplicons of 668 bp were cloned in pDrive vector and
sequenced from both sides followed by sequence and phylogenetic analysis.
Results
RT-PCR based findings confirmed the positivity of CoV in 16.92% (11/65) of cattle,
22.73% (5/22) of Buffalo and 9.38% (6/64) of caprine samples, respectively. Overall
percent positivity for the CoV infection was 14.56% (22/151). Cloning and sequencing
of representative CoV positive samples from cattle for N gene and replicase gene
(ORF1b) revealed that the study strains are quite distinct and remain outside the
major clade comprising of different species of CoV in the phylogram.
Conclusions
The findings of study affirms that CoV infection exists in large and small ruminant
population in the country and additionally the strains of CoVs circulating in Indian
livestock population are genetically diverse. More surveillance studies are needed to
know the exact status of CoV infections in farm animals.

CRISPR/Cas9- Mediated Genomic Deletion of the HERC-5 gene in B95a (Marmoset

B-lymphocyte cells) to limit the PPRV replication

Shikha Saxenaa , Amit Ranjan Sahua, Aruna Pandeya, Sajad Ahmad Wania, Piyali
Mondala, Raja Ishaq Nabi Khana, Waseem Akram Mallaa, Sonalika Mahajana, Bina
Mishrab, Ashok K Tiwaric, Bishnu Prasad Mishraa, Raj Kumar Singha, Deepak Kumara*,
Ravi Kumar Gandhamd*

a Division of Veterinary Biotechnology, ICAR-IVRI, Izatnagar, Bareilly, UP-243122; b

Division of Biological Products, ICAR-IVRI, Izatnagar, Bareilly, UP-243122; c Division
Standardization, ICAR-IVRI, Izatnagar, Bareilly, UP-243122; d National Institute of
Animal Biotechnology, Hyderabad, AP- 500075

[email protected]

Background
Peste des petits ruminants (PPR), a highly contagious disease affecting goats and
sheep. Sungri/96 vaccine strain is widely used for mass vaccination programs in India.
Among the differentially expressed highly connected genes in goats and sheep, HERC-
5 was found to be highly upregulated antiviral gene, identified previously by
transcriptome analysis. B95a cells express high level of SLAM and extensively used for
the infection of PPRV. In this study the role of the HERC-5 gene in isgylation has been
studied by knockout with CRISPR-cas9 in B95a cells for the PPRV replication.
Materials and Methods
In this study to knockout the HERC-5 gene by CRISPR/cas9, three gRNA (guide RNA)
were designed by four different softwares. Three common gRNA in all the softwares
were chosen for the study. The activity of the gRNA was checked by in-vitro cleavage
assay. One gRNA was found to be best among three and cloned in GeneArt
CRISPR.Ofp vector. The cloning was confirmed by PCR, and Sanger sequencing. The
(GeneArtCRISPR.HERC5) was transfected in B95a cells to study the gene knockout and
PPRV replication.
Results
Among the three gRNA for the HERC-5 gene the one in exon 1 was found to be the
best by in-vitro cleavage assay. The first gRNA1 of HERC-5 gene was cloned in
GeneArtCRISPR.Ofp and the genomic DNA of the cloned product was analysed by
sequencing. By transfection of the HERC5 clone, the OFP expression was studied by
FACS. The gene editing efficiency in B95a cells was confirmed cleavage assay and
sequencing. After gene editing the B95a cells studied for PPRV replication by titration
and Real time PCR.
Conclusion
The study demonstrates the role of HERC5 gene as an antiviral agent. Our findings
contribute to a growing body of evidence suggesting that HERC5 is a novel host
restriction factor.