Blood-borne viruses

Prepared by :
Ahmed Abd El-Aziz Mahmoud
Under supervision of :
Dr.Mohamed Ahmed El-Mokhtar
Hepadnavirus
(HBV)
 HBV (hepatotropic)

Family Hepadnaviridae

Envelope Enveloped

NA Partially dsDNA (rcDNA)

Genotypes 8 (A to H)

Transmission Parenteral

sexual

Diseases Acute and chronic hepatitis

Cirrhosis
HCC (2/3 cases)

Diagnosis LFTS
PCR
HBV Markers
Structure
Genome
Mode of transmission
 HBV infection
sequelae

35% Acute
65% subclinical
hepatitis

Chronic carrier (5-10%)

Recovery (90%) Fulminant hepatitis (1%)
HBsAg for >6 months

Asymptomatic Complicated
Death
Carrier state 10%

Chronic active
hepatitis

Cirrhosis

HCC(2/3 cases)
Replication
cycle
 (10000:1)

Virus secretion

Virus entry by
endocytosis

Recycling
Nuclear import

Repair translation (DNA dependent DNA

mechanism
transcription

replication) (+)STRAND
RT
(RNA dependent DNA

Encapsidation
replication) (-) Strand
Immune-evasion in HBV chronic infection

HBeAg excreted to blood (extracellular)

Subviral particles ( immune decoy) :
exhausted Humoral immunity B-Cell mediated IR through Th2 pathway
So suppress Th1 /CMI pathway
Directly inhibit Toll-like receptor (tolerance)
Relaxed circular DNA (rcDNA) has 4 lesions that repaired by the host DNA
repair processes to convert it to Covalently closed circular DNA (cccDNA)

1) HBV-DNA Pol adduct 3) RNA primer/flap

It block repair of both strands (steric

hindrance)
so Should be removed by
deproteination to generate
deproteinated rcDNA to initiate
repair process

2) 10 nucleotide DNA flap 4) ssDNA gap

 1) Removal of Protein adduct (deproteination)
5′-tyrosylphosphodiester Self release: Topoisomerase 1 dp-rcDNA type A
bond in (-) strand
T y rosyl- DNA
P2 (
TDPs:TD te rase)
Serine P
rotease
o d ie s
Phosph

Endonuclease :FEN-1

dp-rcDNA type A dp-rcDNA type C

dp-rcDNA type B

Without Pol Adduct Without Pol Adduct With a short peptide Adduct
With DNA flap (r sequence ) Without DNA flap With DNA flap
 3)Sealing the nick
2) Removal of DNA flap

LIG-1 , LIG-3
FEN-1 (Flap
Endonuclease) Ligases

Repaired (-) strand

dp-rcDNA type B
dp-rcDNA type A&C

4) Filling the ssDNA gap

POLδ, POLκ, POLι, and POLη

cccDNA RFC(clamp loader), PCNA(sliding
clamp)

LIG-1 , LIG-3 FEN-1 (Flap

Ligases Endonuclease)

Repaired (+) strand 6)Sealing the nick 5) Removal of RNA flap

Reverse
transcription
Reverse transcriptase ( pg-RNA to rc-DNA)

Structure: 4 domains Terminal protein domain

has tyrosyl residue that
Reverse bind covalently to GAA
transcriptase(RDDP/ through priming initiation
DDDP activities) and polymerization

Spacer domain

Rnase H domain
Degrade pg-RNA
during synthesis of
minus strand
Steps of pg-RNA reverse transcription
Step 1: Primer synthesis
1) Epsilon hairpin has 5`-UUC-3` which is the template for 5`-GAA-3` nucleotide
that is covalently bounded to the tyrosine residue of TP and act as a primer for
synthesis initiation.
Step2: Translocation and binding to DR-1(direct repeat 1 sequence ) at 3` end of pg-RNA
and elongation of minus strand.

Step3: ) Synthesis of (-) DNA strand with concurrent degradation of pg-RNA by RNase H domain
Step4: Circularization of the (-) DNA strand through covalent attachment of the polymerase at its
5`end and preservation of the RNA primer (DR1) from the 5`end of the pgRNA.

Step5: Hybridization of the DR1 RNA primer with DR2 at the 5` end of the (-)DNA strand and
extension for (+) DNA strand Synthesis.
Mutations in HBV genome
 Diagnosis

Viral DNA Detection By

LFTs HBV markers
PCR & DNA probe

Indicate viremia
3 Viral Ags Used for monitoring
ALT/AST
& their Abs by ELISA/RIA Antiviral drugs effect
HBV Markers interpretation
Test Acute infection Resolved acute High Low vaccinated Window phase
infection replication replication
chronic chronic
infection infection

HBsAg + - + + - -

HBeAg (transmissibility + - + - - -
factor /infectivity
factor)

Anti-HBs - + - - + -

Anti-HBe - + - + - -

Anti-HBc IgM + - + - - +

Anti-HBc IgG - + + + - -

AST/ALT +++ Normal +++ Normal Normal

Serological pattern of HBV acute infection
Serological pattern of HBV chronic infection
Phase of HBV chronic infection

High replication
High DNA level
Low replication High inflammation (ALT)
Low DNA level HBeAb +ve
Low inflammation (ALT) TTT
HBeAb +ve
High replication
High DNA level
Low inflammation (ALT)
HBeAg +ve
High replication
High DNA level
High inflammation (ALT)
HBeAg +ve
TTT
Prevention and control
1) General measures :

• Avoid close contact with blood and body fluids

• Supervision of blood transfusion

• Use disposable syringe

2) Pre-exposure prophylaxis :
Recombivax 3 doses IM in deltoid region at 0,1,6 m` intervals
(dose=0.5 <10 years//1mL >10 years )

3)Post-exposure prophylaxis :
human Ig followed by active vaccination
Treatment
• pegylated IFN-Alpha 2a (pegasys /SC/Q.WK) in
chronic hepatitis

• Lamivudine (Q.D )and Adefovir (nucleoside

analogues that inhibit viral DNA polymerase)

• Infants born to HBsAg +ve mother (single dose

HBIG 0.5 mL within 12hr of birth) followed by
active immunization with vaccine
Flavivirus
(HGV)
 HGV (lymphotropic)
Family Flaviviridae

Envelope Enveloped

NA +ve sense
ssRNA

Genotypes 7 (genotype 1)

Transmission Parenteral

sexual

Diseases 1) Chronic GBV-C Viremia

Among healthy blood donor (1-4%)

2) co-infection with other parentally transmitted

Viruses

3) Suspected to cause Post-transfusion hepatitis (NANE)

Diagnosis PCR
Ab detection (anti-E1/anti-E2)
HGV(HPgV / GBV-C) [lymphotropic]

Single ORF (2 structural /5 nonstructural protein )

Hepatitis G may be associated with favorable outcomes in
patients with HIV infection.
Circoviridae
(TTV/SEN-V)
 TTV (transfusion transmitted virus) SENV (named after the initials the 1 st
patient)
Family Circoviridae Circoviridae

Envelope Non-enveloped Non-enveloped

NA -ve sense ccc -ve sense Circular
ssDNA ssDNA virus

Genotypes 16( 1a/1b) 8 (A to H)

Transmission Parenteral Parenteral

Enteric sexual

Sexual

Diseases 1)Suspected to cause Post-transfusion 1)Suspected to cause Post-transfusion

hepatitis (NANE) hepatitis (NANE)

2)Chronic viremia in apparently healthy 2)Chronic SEN-V viremia

blood donor (infection without clinical
symptoms)

Diagnosis PCR PCR

Ab detection (anti-TTV /IgM anti-TTV)

85% of blood donor TTV +ve(genotype 1,2

mainly)
Thank you