1. IMPORTANCE OF HCV GENOTYPING
AND
MULTIPLEX PCR FOR GENOTYPING
By,
Ushanandini Mohanraj

2. FACTS:-
ONLY 20% OF PEOPLE INFECTED WITH HEPATITIS C VIRUS
WILL COMPLETELY RECOVER
HCV IS 10 TIMES MORE CONTAGIOUS THAN HIV WHEN
COMPARING DIRECT BLOOD-TO-BLOOD CONTACT
15 MILLION PEOPLE IN INDIA ARE AFFECTED BY HEPATITIS C
SILENT DISEASE
MORE THAN 3 LAKHS FOR CURRENT TREATMENT REGIMENT OF
24- 48 WEEKS WITH SIDE AFFECTS

3. WHY GENOTYPE HEPATITIS C VIRUS?
Genotype HCV Treatment Recommendations by
AASLD
1 Harvoni for 12 weeks
2 Sovaldi + ribavirin for 12 weeks
3 Sovaldi + ribavirin for 24 weeks
4 Harvoni for 12 weeks / Sovaldi + ribavirin for
24 weeks
5 Sovaldi + ribavirin + PEG for 12 weeks
6 Harvoni for 12 weeks

4. Molecular methods:-
1] Innolipa
2] Restriction Fragment length
polymorphism
3] Phylogenetic analysis and sequencing
Serological method:-
1] Use of genotype specific antibodies
GENOTYPING METHODS

5. INNO LIPA
EXCEPT GENOTYPE 6 GOOD FOR GENOTYPING
AT SUBTYPE LEVEL ONLY 1b AND 3b DISTINGUISHED ACCURATELY

6. PHYLOGENETIC ANALYSIS AND SEQUENCING:-
THREE STEPS:-

10. - IS GENOTYPING OF HEPATITIS C VIRUS
BASED ON PHYLOGENETIC ANALYSIS
OF 5’UTR REGION ACCURATE?
- IS THERE A NEED TO
ANALYSE OTHER REGIONS
OF HCV GENOME?

11. NS5B REGION:-
Sofosbuvir, an antiviral drug targeting NS5B,
approved by the FDA in December 2013.
1.1 LAKHS FOR 24 WEEK TREATMENT
Dasabuvir , Non-nucleoside NS5B polymerase inhibitor ,
Approved by FDA in December 2014
1000 BASES

12. CORE REGION:-
500 TO 600 BASES
HCV core protein is a potential
modulator of the host immune system

13. NS5B CORE

14. 1]Specificity
2]Accurate results
1] sequencing is time consuming
and a costly affair
2]sequencing of amplified DNA
does not usually identify
mixed infections in patients.

15. IDEAL GENOTYPING METHOD:-
1] ACCURATE AND RELIABLE
2] CHEAP
3] FASTER RESULTS

16. “GENOTYPING USING MULTIPLEX PCR”
Key features:-
1] results with just
electrophoresis
2] No need of sequencing
3] less cost and time

17. CHALLENGES:-
-- DECIDE WHICH REGION TO FOCUS
-- TEST EFFICIENCY
-- DESIGN SPECIFIC PRIMERS
-- NEED FOR GENOTYPING STUDIES

18. IS GENOTYPING USEFUL FOR ONLY
TREATMENT RECOMMEDATIONS?

19. -- The geographic distribution and diversity of HCV
genotypes
-- Tracing the source of an HCV outbreak in a given
population.
-- Potential Role of HCV heterogeneity and genotypes in
mother-to-infant Transmission of HCV
-- The role of HCV genotypes in the progression of liver
disease is one of the most controversial areas of HCV
research
WHATS’ MORE TO THE GENOTYPING RESEARCH?