1. In The Name of God

2. Diagnosis of HBV & HCV
Dr Mostafa Javanian

3. The laboratory finding were as follow:
A HCW Presented with icter and fever, one month after Needle stick with a known case of chronic HBV infection.
The laboratory finding were as follow:
Medical university of Babol

4. ALT=4100 IU/l AST=3600 IU/l ALK =500 IU/l Bill T: 10mg/dl D: 6mg/dl
HBS Ag :positive
Anti HBC IgM : positive
HBe Ag : positive
HBs Ab : negative
Anti HCV : negative
Anti HAV IgM : negative
HIV ELISA : negative
Medical university of Babol

5. HBV Infection
Medical university of Babol

6. Laboratory Tests for HBV
Serology:
Many tests available – most common tests are Enzyme Immunoassays (EIAs, MEIAs)
First tests available in 1972
For every rule, there is an exception/caveat
No single test tells you everything
Molecular:
HBV DNA (quantitative)
HBV genotyping
HBV resistance testing
Medical university of Babol

7. Hepatitis B – Laboratory Tests
Serologic markers:
1) HBsAg (Hepatitis B surface antigen):
if positive, person is infectious
Sensitivity = 0.15 ng/ml
Specificity = 99.5%
2) Anti-HBs (Antibody to HBV surface antigen):
indicates immunity to HBV and protection from disease
Protective level is >10 IU/ml
Medical university of Babol

8. Hepatitis B – Laboratory Tests
Serologic markers:
3) Anti - HBc (Antibody to HBV core antigen):
Total - indicates past or active infection; present whether person is immune or chronic carrier
Specificity = 99.8% to 99.9%
IgM - early indicator of acute infection
No antigen test
Medical university of Babol

9. Hepatitis B – Laboratory Tests
Serologic markers:
4) HBeAg (Hepatitis Be antigen):
indicates person is highly infectious
Selecting patients for therapy
5) Anti-HBe (Antibody to HBVe antigen):
prognostic for resolution of infection; less infectious; spontaneous seroconversion in 10 to 20% of healthy adults per year
Medical university of Babol

10. Hepatitis B – Laboratory Tests
Serologic markers – caveats:
Persistent HBsAg for >6 mos = chronic infection
HBsAg and anti-HBs may co-exist in some of chronically infected individuals; likely due to mutations in the “a” determinant of the S gene
Surface antigen escape mutants described in infants infected with HBV after HBIG + vaccination and in Liver transplants after prolonged HBIG
Anti-HBc IgM may persist for up to 2 years in 20%; chronically infected individuals may have low titres which rise during acute flares
Medical university of Babol

11. Hepatitis B – Laboratory Tests
Serologic markers – caveats:
Precore or HBeAg negative mutants:
Due to mutation in precore (abolishes HBeAg production) or core promoter region (down-regulates HBeAg production)
No effect on viral replication (may be enhanced)
More difficult to treat; greater risk of cirrhosis
Co-infection with HCV may suppress both HBeAg and HBsAg
Medical university of Babol

12. Hepatitis B – Laboratory Tests
Serologic markers – caveats:
Isolated HBcAb may be due to:
Remote infection (immune or chronic carrier)
“Window” period between HBsAg and HBsAb
Co-infection with HCV
False positive test result – HBcAb is marker most prone to false positives
HBV DNA may help sort this out
Medical university of Babol

13. Medical university of Babol

14. Disease Phases in Chronic HBV Infection
HBsAg
HBeAg
Anti-HBe
ALT
HBV DNA range
Immune Tolerant + -
Normal
>8 log IU/mL
Immune Clearance
Normal or elevated
3-8 log IU/mL
Inactive Disease
<3 log IU/mL
HBeAg-negative Chronic HBV
Medical university of Babol

15. HEPATITIS C VIRUS
Medical university of Babol

16. Laboratory Tests for HCV
Serology:
Detection of anti-HCV antibodies
Serologic test available since 1990
Molecular:
HCV RNA detection
Determination of HCV genotype
Viral load determination
The diagnosis of HCV infection is based on the detection of specific antibodies and the viral RNA genome.
Medical university of Babol

17. Laboratory Tests for HCV
Serology:
Screening:
3rd generation EIAs measure antibodies directed against recombinant peptides NS4, core, NS3, and NS5 proteins
Sensitivity = 97%
Detects antibodies within 6 to 8 weeks
No HCV IgM test available
Confirmatory/supplementary:
RIBA, Second EIA, HCV RNA
The diagnosis of HCV infection is based on the detection of specific antibodies and the viral RNA genome.
Medical university of Babol

18. Laboratory Tests for HCV
Molecular:
Both qualitative and quantitative HCV RNA assays available
Used for treatment monitoring (and in some circumstances for confirmation of positive or indeterminate serology)
HCV RNA is detectable 2 to 14 days after an exposure
The diagnosis of HCV infection is based on the detection of specific antibodies and the viral RNA genome.
Medical university of Babol

19. Genotyping Used for: Methods include:
Detection of mutations that confer resistance to antiviral agents
Genotyping of isolates for epidemiological purposes; categorizes patient isolates into 8 different HBV genotypes (A to H) and 6 different HCV genotypes (1 to 6 with 24 subtypes)
Methods include:
Sequencing
Hybridization (Line Probe Assay, Trugene Assay)
Medical university of Babol

20. Diagnostics in Viral Hepatitis: Summary
Serology remains the cornerstone for diagnosis and screening
NAAT is critical to patient management
Of the many NAAT tests available, PCR, bDNA and TMA remain most popular
Sensitivity and dynamic range varies between assays
Standardization allows (to some degree) interchangeability of the results with different assays
Resistance/Genotyping requires amplification first
Increasing role in making treatment decisions as more drugs become available for HBV
Medical university of Babol

21. با تشکر فراوان از توجه شما