Pandemic SARS Coronavirus-2 Infections in Humans- COVID-19

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N. MUSTAFA, H. ZAHOOR, F. M. MAJOO
Pandemic SARS Coronavirus-2 Infections in Humans-
COVID-19
Nasir MUSTAFA*, Hina ZAHOOR**, Fuzail M. MAJOO***
Abstract
The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) first broke out in Wuhan
(China) and subsequently spread worldwide. Coronaviruses (CoVs) primarily cause zoonotic
infections in birds and mammals however, in the last few decades have shown to be capable of
infecting humans as well. The outbreak of severe acute respiratory syndrome (SARS) in 2003
and more recently, Middle-East respiratory syndrome, (MERS) has demonstrated the lethality
of CoVs when they cross the species barrier and infect humans. Coronavirus (CoV) is a large
family of viruses that cause afflictions ranging from the common cold to more severe
pathologies such as Middle East Respiratory Syndrome (MERS-CoV) and Severe Acute
Respiratory Syndrome (SARS-CoV). A novel coronavirus (nCoV) is a new strain that has now
been identified in humans. The recognition of a new coronavirus identified in December 2019,
named CoVID-19 are common for coronavirus researchers. Detailed investigations found that
SARS Coronavirus-2 was initially transmitted from civets to humans and MERS-CoV from
dromedary camels to humans. Advances in biology have resulted in a greater understanding of
coronavirus, including them to adapt to new environments, trans-species infection and the
emergence of new subtypes. New tools of cell and molecular biology have led to an increased
understanding of intracellular replication and viral cell biology. Along with the advent of
reverse genetic approaches in the past five years; it is now possible to begin to define the
Derleme Makale (Review Article)
Geliş / Received: 10.03.2020 & Kabul / Accepted: 28.03.2020
* Assist. Prof. Dr., Istanbul Gelisim University, School of Health Sciences, Department of
Nursing (English), Istanbul, Turkey, E-mail: nasirmustafaspan@gmail.com
ORCID ID https://orcid.org/0000-0002-5821-9297
** Lecturer, Istanbul Gelisim University, School of Health Sciences, Department of Social Work
(English), Istanbul, Turkey, E-mail: hinazahoor79@gmail.com
ORCID ID https://orcid.org/0000-0003-2322-5678
*** Lecturer, Istanbul Gelisim University, School of Health Sciences, Department of Nursing
(English), Istanbul, Turkey, E-mail: fmajoo@gelisim.edu.tr ORCID ID https://orcid.org/0000-
0002-4413-8795

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determinants of viral replication, trans-species adaptation, and human disease. The most
progress has been made on SARS-CoV 2, highlighting specific structural requirements for its
functions in the CoV life cycle as well as mechanisms behind its pathogenesis. In this review, we
will provide a through insight to the life cycle of CoV, its genetics, replication process and
reverse genetic applications to SCoV along with advances in its research. This review aims to
establish the current knowledge on CoV-2 by highlighting the recent progress that has been
made and comparing it to previous knowledge. We also conclude with a brief discussion on
practices to decrease risk factors for transmission and treatment options.
Keywords: Human coronavirus, SARS coronavirus-2, infections in humans-COVID-19.
İnsan-COVID-19'da Pandemik SARS Coronavirus-2 Enfeksiyonları
Öz
İlk olarak Wuhan'da (Çin) patlak veren Şiddetli Akut Solunum Sendromu Koronavirüs 2 (SARS-
CoV-2), sonrasında dünyaya yayılmıştır. Koronavirüsler (CoVs) başlarda kuşlarda ve
memelilerde zoonotik enfeksiyonlara neden olurken, son birkaç yılda insanları da enfekte
edebildiği gösterilmiştir. 2003 yılında Şiddetli Akut Solunum Sendromu'nun (SARS) ve son
zamanlarda Orta Doğu Solunum Sendromunun (MERS) patlak vermesi, CoV'lerin tür bariyerini
geçip insanları enfekte ettiklerinde ölümcül olduklarını göstermiştir. Koronavirüs (CoV), soğuk
algınlığından Orta Doğu Solunum Sendromu (MERS-CoV) ve Şiddetli Akut Solunum Sendromu
(SARS-CoV) gibi daha şiddetli patolojilere kadar çeşitli rahatsızlıklara neden olan büyük bir
virüs ailesidir. Yeni koronavirüs (nCoV), insanlarda daha önce tanımlanmamış yeni bir suş
türüdür. Aralık 2019’da tanımlanan yeni koronavirüs, araştırmacılar tarafından genel olarak
CoVID-19 olarak adlandırılmıştır. Yapılan ayrıntılı araştırmalarda, SARS Coronavirus-2'nin
başlangıçta misk kedilerinden, MERS-CoV'un ise tek hörgüçlü develerden insanlara bulaştığı
bulunmuştur. Biyoloji alanındaki gelişmeler koranavirüsün, yeni ortamlara adaptasyonu, trans-
tür enfeksiyonu ve yeni alt tiplerinin ortaya çıkması da dahil olmak üzere daha iyi anlaşılmasını
sağlamıştır. Hücre ve moleküler biyolojide kullanılan yeni araçlar, hücre içi replikasyon ve viral
hücre biyolojisinin daha iyi anlaşılmasını sağlamıştır. Son beş yılda ters genetik yaklaşımların
ortaya çıkmasıyla birlikte; artık viral replikasyonun, trans-tür adaptasyonunun ve insan
hastalığının belirleyicilerini tanımlamaya başlamak mümkündür. Bu derlemede, CoV'nin yaşam
döngüsü, genetiği, replikasyon süreci ve SCoV'ye ters genetik uygulamalarının araştırılması ile
ilgili ilerlemelerle ilgili bilgi verilecektir. Bu derleme, yakın zamanda kaydedilen ilerlemeyi
vurgulayarak ve önceki bilgilerle karşılaştırarak CoV-2 hakkındaki mevcut bilgileri oluşturmayı
amaçlamaktadır. Ayrıca, bulaşma ve tedavi seçenekleri için risk faktörlerini azaltmaya yönelik
uygulamalar hakkında kısa bir tartışma ele alınmıştır.

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Anahtar Kelimeler: İnsan koronavirüsü, SARS koronavirüs-2, insan-COVID-19
enfeksiyonları.
Introduction
Life Cycle of Coronavirus
Advances in biology have resulted in a greater understanding of coronavirus, including
structure genome and nucleic acid sequence. The Coronavirus genome is a linear RNA
particle consisting of a spike, an enveloped layer and a nucleic acid sequence with a
single strand of mRNA1. The genome is composed of the nucleocapsid protein N and
contain an RNA nucleocapsid helical structure. The S protein is the evidence of
pathogenesis as observed in mice. It contains species specificity, range, cell tropism and
a disease marker2. On the host cell area, virus replication initiates when the S protein
conforms to particular receptors. Morphologically the genome of coronaviruses is
single-stranded, linear mRNA molecules with polarity and diversity of lengths ranging
from 28 to 32 kb3. Aminopeptidase N is the receptor in human coronavirus HCoV-
229E. The precise mechanisms of entry and uncoating have not been defined, but likely
occur by either fusion from viral and host cell membrane or through endocytic vesicles.
The next discrete stage in the life cycle is translation and proteolytic processing of viral
replicase proteins from the input genome RNA, followed by formation of cytoplasmic
replication complexes in association with cellular membranes4. Mechanisms by which
replication products are delivered to sites of assembly remain to be determined. It has
been shown that subpopulations of replicase proteins and the structural nucleocapsid
(N), translocate from replication complexes to sites of assembly and may mediate the
process in association with cellular membrane protein pathways5. The endoplasmic
reticulum Golgi intermediate compartment (ERGIC) viral congregation occurs both
rapidly and physically distinct from viral replication structures. The replication
mechanisms involved in the sites of influence, remain to be determined. It has been
observed that replication protein and nucleocapsid N, alter from replication
organization place together and arbitrate this system in combination with cell protein
film pathways6. The genome RNA N, layer protein E and protein M adhere into the
lumen of the ER/Golgi virosomes, which results in the budding and formation of
particles7.

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Viral Replication
The 5′ replicase superiority of the RNA genome divides into co-amino terminal
replicase polyproteins. For a continuous reproductive life cycle; coronavirus infection
replication needs constant replicase quality of translation8. Replicase proteins arbitrate
through positive and negative strand, genomic and subgenomic RNA production, as a
result of capping, polyadenylation, RNA recoiling, arrangement change during viral
RNA processing, and alternating transcription and transcription attenuation9. With the
advent of SARS, more extensive bioinformatics analyses have resulted in predictions of
several additional functions involved in RNA processing, including methyltransferase
and exonuclease activities10.
Replicase Protein Expression
Coronaviruses activity incorporate both picornavirus 3C-like proteinase and papain-
like proteinase (PLP) which are needed for encoding inside the replicase polyproteins,
which equally mediate both trans and cis cleavage trial11. There are sites for replicase
polyproteins by amino acid sequences and the region of the replicase which experience
the majority variation, suggestive for encoding and adornment functions or specific
host jobs which are responsible to demands for quick additional alteration12.
Figure 1: (A) Ultrastructural morphology exhibited by coronaviruses.
Figure 1: (B) 3D medical Images, depicting the shape of coronavirus.
Figure 1: (C) Cross-sectional view shows the major elements.

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Figure 2: The infection cycle of a coronavirus13
Expression of Structure
Only the 5′ most replicase gene is translated from the input positive-strand genome
RNA. The genome contains multiple genes for the known structural proteins S, E, M,
and N, as well as other genes for expression of proteins that have been labeled as
“nonstructural” or “accessory” because they have been presumed not be required for
replication. Moreover, they are thought not to be incorporated into virions. MHV
encodes six of these genes, while SCoV encodes possibly up to 11 structural and
accessory genes, which are expressed from subgenomic mRNAs14. Furthermore, the

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deletion of a single extra character in individual isolates of SCoV, suggested that this
character is concerned in the alteration for replication and transmission in humans15.
Genetics of Coronavirus
With the help of technological advances in the field of genetics, researchers have been
able to learn about the pathogenesis and replication of CoV. The genetics of coronavirus
replication and pathogenesis have largely been studied using natural variants, host
range mutants, passaged virus, and mutagenized viruses selected for temperature
sensitivity and specific phenotypes. Classical complementation of functions made it
possible to define at least eight genetic groups for MHV, with most of the
complementation groups localized to the replicase gene16. Research reveals that the
major marker of tropism and pathogenesis is the S protein. Furthermore, the genetic
segment in replicase may be responsible for the distinctive activities of coronavirus.
Specific modification and groundbreaking range have been limited to studies of the 3′
10 kb of the MHV genome and restrict the selection of viable recombinants. The
establishment of infectious clone reverse genetic strategies for the corona viruses
transmissible gastroenteritis virus (TGEV); a better understanding of its genome has
been made possible by utilizing "infectious clone" reverse genetic strategies17.
Gastroenteritis Coronavirus Transmissible
HCoV-229E, IBV, and MHV has made it theoretically possible to study the genetics of
the entire genome and all of the structural, accessory, and replicase genes. Approaches
to “infectious cloning” have included full-length cDNA clones of TGEV genome in
bacterial artificial chromosomes. The in vitro assembly approach was developed to
overcome the challenge of full-length cDNA cloning of the TGEV and MHV genomes,
which contained “toxic” regions in the replicase gene, resulting in unstable or toxic
clones in E. coli18. In combination with biochemical and cell imaging approaches, it is
also possible to study highly defective or lethal mutations in electroporated cells in
order to define critical determinants of replication. The in vitro assembly approach has
been used to introduce marker mutations that are silent for replication in culture. Thus,
direct reverse genetic studies of the critical replicase gene functions can be performed
using in vitro assembly of infectious clones19.

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Advances Research About SCoV
The initial research of coronaviruses that had been conducted by the National Institutes
of Health, lead to the quick advancement in the recognition and study of SCoV as the
etiologic instrument of SARS, in addition to the ability of the virus to viably grow in
cultures20. Corresponding knowledge of CoV was viable thanks to the swift detection of
ACE II as the receptor for SARS; which verified the great importance of research of
coronavirus subtypes. Likewise, replicase feature expression, handling and anticipated
function have made it possible for therapeutic approaches and identification of genome
structure studies21.
Reverse Genetics Studies Application
Robust tissue culture systems and emerging animal models, create the potential to
rapidly answer questions concerning: (1) determinants of virus growth in culture; (2)
potential mechanisms of transspecies adaptation; (3) sensitivity to and escape from
biochemical and immune interference with replication; (4) determinants of virulence
and pathogenesis; (5) mechanisms of genome recombination and mutation; (6)
functions of and requirements for replicase, structural, and accessory proteins; and (7)
development of stably attenuated viruses for use as seed stocks for inactivated vaccine
or testing as live-attenuated vaccines22. While the Coronavirus genome and its protein
makeup demonstrate great tolerance of modification, all sequences of quality-based
insertion, deletion or mutagenesis, have so far answered to have viruses damage in
replication, pathogenesis, or both. Large numbers of changes in MHV and other species
of coronaviruses are observed in ScoV. This technique could be an attempt for likely
attenuation in SCoV culture and animal models. Finally, it is essential to generate
strains of SCoV that are weakened and stabilized against reversion and recombination;
to discover factors for replication and pathogenesis determinants and construction of
virus. Such weakened modifications would offer supplementary precautions in
determining SCoV development, biology, disease, treatment, and prevention23.
Coronaviruses in Human
Vaccines are components that are not therapeutic but protective. Seven strains of
human coronaviruses have been identified (i) Human coronavirus HCoV-229E (ii)
Human coronavirus HCoV-OC43 (iii) Severe acute respiratory syndrome

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coronavirus SARS-CoV (iv) Human coronavirus HCoV-NL63, New Haven coronavirus
(v) Human coronavirus HKU1 (vi) Middle East respiratory syndrome
coronavirus MERS-CoV, previously known as novel coronavirus 2012 and HCoV-EMC.
(vii) Severe acute respiratory disorder coronavirus SARS-CoV-2, previously known as
nCoV-2019 or "novel coronavirus 2019". The coronaviruses HCoV-229E, - NL63, -
OC43, and - HKU1 frequently move in the human population and cause respiratory
infections in adults and children widely24.
Table 1: Characteristics of patients who have been infected with SARS-CoV-2,
MERS-CoV, and SARS-CoV25
Conclusion
Coronavirus is highly pathogenic in nature. HCoVs causes serious intense respiratory
disorder. For SARS-CoV and MERS-CoV, animal reservoirs were the source for the
worldwide outbreak with shocking morbidity and mortality26. The recognition of a new
coronavirus identified in December 2019, named CoVID-19 are common for
coronavirus researchers. CoVID-19 was documented in Wuhan, China, and has caused

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severe infection and death globally. Some healthcare professionals have theorized that
2019-nCoV could progress to a low pathogenic yet highly infectious coronavirus, which
may reoccur annually every winter such as the annual influenza virus. The critical
expansion and outlook of this epidemic at the time of this writing is unknown as the
virus is rapidly differentiating27. Coronaviruses can be classified into four genus: alpha,
beta, gamma, and delta, in which alpha and beta CoVs transmit the disease to humans.
Four HCoVs (HCoV 229E, OC43, NL63 and HKU1) are globally prevalent, all wide-
ranging as evidenced by 30% of upper respiratory tract infections in adults28.
Coronaviruses are ecologically dissimilar with the most excellent diversity seen in bats,
signifying that they are the main source in the production of large quantities of the
virus. Warm-blooded animals are considered as an intermediate host; assisting in
recombination and mutation extension of genetic variety. Normal symptoms of SARS,
incorporate with pyrexia, coughing, dyspnea, and diarrhea. Thirty-two percent of
infected patients require mechanical ventilation while 10% result in death29. Increased
fatality rates are observed in patients with medical comorbidities. Human to human
transmission was evident chiefly in medical settings. The Nosocomial extension might
be explained by basic virology. The major human receptor for the SARS S glycoprotein,
the angiotensin-converting catalyst II (ACEII), is found chiefly in the lower respiratory
tract, as opposed in the upper reparatory passage. MERS, SARS and the Wuhan
coronavirus appear to cause more severe disease in older people, though uncertainty
remains around the latest outbreak. Of the cases of Wuhan coronavirus reported so far,
none are yet confirmed to be among children and old age group. Receptor sharing may
represent upper respiratory tract signs and symptoms that top viral flaking occurring
late into the tenth day of infection when the patient is hospitalized30. SARS care often
requires vaporized treatment protocols such as intubation, which may promote and
increase nosocomial activity. The surface spike glycoprotein is dangerous for the
adhering of host cell receptors and considered as a key determinant of host run
restriction. HCoV has remained relatively low key due to their weak phenotypes in
humans31. This changed in 2002, when cases of severe atypical pneumonia were
described in Guangdong Province, China, causing worldwide concern as the disease
spread via international travel to more than 202 countries. The disease got known as
Severe Acute Respiratory Syndrome (SARS), and a beta HCoV, named SARS-CoV, was
recognized as the causative factor. Since the new cases shared account of human-

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animal contact at live game markets, zoonotic transmission of the virus was definitively
supported. Palm civets and raccoon hounds were originally considered to be the animal
reservoir. A few vital events occurred in select populations such as the well described
mini flare-up in the Hotel Metropole in Hong Kong, where infected subjects traveled
and spread SARS globally32. Another incident occurred at the Amoy Gardens housing
complex where more than 310 people were infected, giving proof that air transmission
of SARS-CoV may take place from time to time. About two decades later, the
mechanism linked with the transmission of SARS-CoV, ranging from self-limited
animal to human transmission to human–human transmission instance, remain
unsuccessfully understood. Currently, while there is no treatment, researchers are at
work. As is the case for most viral infections, symptoms usually subside independently.
Health care providers suggest seeking professional care early. In the event that signs,
and symptoms feel more severe than a standard cold, patients are recommended to
seek immediate medical care. Physician can reduce side effects by prescribing pain or
fever medication33.
Table 2: A comparison of diagnostic criteria based on symptoms and travel34

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A room humidifier or a hot shower can help with a sore throat. Increased fluids
consumption, rest and increasing the time of rest, all show to improve recovery35. The
Wuhan coronavirus mortality rate is greater than SARS and MERS, yet still similar to
the 1918 Spanish influenza pandemic. Researchers believe the mortality rate is likely to
be less because of an "iceberg" of milder cases are yet to be found. This highlights that
novel viruses multiply far more rapidly through a population. Presently, a vaccine to
safeguard against this family of viruses does not exist. Trials for a MERS vaccine are in
progress. The US National Institutes of Health is working on a vaccine against the new
species of virus. It will be a long time before clinical trials get in progress and greater
than a year until they are made available. People may be able to reduce the risk of
infection by avoiding people who are sick. Individuals must avoid touching the eyes,
nose and mouth. Most people who become infected experience mild illness and recover,
but it can be more severe for others. Washing the hands often with soap and water and
for at least 20 seconds or cleansing with hand sanitizers may neutralize viruses that
may be on the hands. Maintaining a social distance of at least 1 meter (3 feet) between
yourself and anyone who is coughing or sneezing; as the mist of small liquid droplets
from the nose or mouth may contain the virus. Because hands touch many surfaces and
can pick up viruses; contaminated hands can transfer the virus through the mucous
membrane of the eyes, nose or mouth and lead to pathogenesis in the host. The practice
of positive respiratory hygiene includes covering your mouth and nose with a bent
elbow or tissue when coughing and sneezing, and the immediate disposal of the used
tissue. This practice should be encouraged and enforced in social communities.
Following positive respiratory hygiene, hand washing/cleansing and avoidance of
touching the eyes, nose and mouth, protect populations from viruses such as the cold,
flu and COVID-19. Individuals with a fever, cough and dyspnea must seek early medical
care. Individuals who feel ill should stay home. Individuals should follow advice given
by healthcare providers and staying informed with the latest developments about
COVID-19. Protective measures should be taken for individuals who are in or have
recently visited (past 14 days) areas where COVID-19 is spreading36. For diagnosis,
most testing for COVID-19 is currently done on viral genetic material from nose and
throat swabs using a workhorse tool of molecular biology known as reverse
transcription-polymerase chain reaction (RT-PCR). Labs worldwide have customized
their PCR tests for SARS-CoV-2, using different primers targeting different sections of

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the virus’s genetic sequence. Protective measures currently being applied by countries
to combat the disease include the screening of travelers. The emergence of 2019-nCoV,
has triggered health screenings at airports in multiple countries. The screenings will
likely not be the deciding factor in whether the virus continues to spread globally.
Quarantine means the separation of a person or group of people reasonably believed to
have been exposed to a communicable disease but not yet symptomatic, from others
who have not been exposed, in an effort to prevent the possible spread of the
communicable disease. Currently, a fourteen-day quarantine is observed prior to days
prior to an assessment in individuals suspected to have exposure. If traveling to China,
learn the sign and symptom, maintain social distance of 3 meters and avoid settings
such as live animal marketplace, which is where the latest breakout is thought to have
originated in Wuhan. On December 31, 2019, Chinese authorities detailed groups of
pneumonia cases in Wuhan, China where a large portion of the effected population was
reported to have frequented live game markets. According to the current
epidemiological situation in the world, the Coronavirus disease (COVID-19) outbreak
has caused 509,164 human infections and 23,335 deaths spanning in 202 countries
(WHO update: 27 March 2020). It is different from SARS-CoV in biological,
epidemiological, and clinical highlights. Researchers at the Infectious Diseases Vaccine
Research Center and the National Institute of Allergy, have made advances in the use of
a nucleic acid vaccine as a possible treatment37. During SARS, experts progressed in
acquiring the genome of SARS-CoV to a stage I clinical trial of a DNA vaccine in 20
months and have since packed down that period to 3.5 months for other viral diseases.
For 2019 nCoV, they desire to shift considerably faster, using representative RNA
(mRNA) vaccine innovation38. The international health authorities are now
brainstorming on the search for therapies able to counter the most severe effects of the
disease. In this regard 600mg of hydroxychloroquine have been found to be efficient on
SARS-CoV-2 patients39. In addition, it is also reported that Hydroxychloroquine is
more effective with azithromycin and it causes significant reduction of the viral activity
so it is concluded that combine therapy of Azithromycin with hydroxychloroquine
significantly more efficient for virus elimination40.

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