Comparison of PCR-based genotyping methods for hepatitis B virus.
Hepatitis B virus (HBV) is classified into eight genotypes (A to H). In this study, three genotyping methods were compared for their sensitivity and accuracy, namely PCR-RFLP on the S region, PCR-RFLP on the pre-S region and nested PCR with type specific primers. Sixty HBV samples from infected sera were genotyped. The nested PCR with type specific primers was found to be the most sensitive and produced substantial numbers of co-infections by genotypes B and C. The sensitivities for both PCR-RFLP methods were lower and did not reveal co-infections. Generally, the three methods produced consistent genotyping results in samples infected by single genotypes but for co-infections by genotypes B and C, the two PCR-RFLP methods yielded only single genotypic results. For the cases of single genotypic infections, genotypes ascertained by sequencing were in concordance across all three methods. However, when co-infections occurred, PCR analysis on clones revealed only single genotypic infections.
Lim
C K
CK
School of Arts and Sciences, Monash University Sunway Campus, Selangor Darul Ehsan, Malaysia.
Tan
J T M
JT
Ravichandran
A
A
Chan
Y C
YC
Ton
S H
SH
D004279:DNA, Viral / Q000032:analysis; D005838:Genotype; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity
D004279:DNA, Viral