Genotyping of hepatitis B virus by restriction pattern analysis of a pre-S amplicon.
A method is described for genotyping of hepatitis B virus (HBV), based on the restriction fragment length polymorphism (RFLP) created by Ava2 and Dpn2 action on an amplified segment of the pre-S region. Analysing 51 database sequences by phylogenetic tree construction and RFLP prediction, the method was shown to be capable of detecting all known genotypes (A-F). The method was applied to 99 serum samples from hepatitis B e antigen (HBeAg)-positive chronic carriers, comparing observed agarose gel patterns with the RFLP predicted from the database sequences. In 95 typable samples the following genotypes were observed; 23 A, 20 B, 20 C, 22 D, 5 E and 5 F. Phylogenetic grouping of the 51 database sequences and RFLP genotyping of the 99 patient samples were compared with typing based on S gene analysis, showing disagreement in only one case, a database sequence of ayw subtype which was classified as genotype D by pre-S region and genotype A by S region analysis. This method should be useful for epidemiological investigations and for studying the potential influence of genotype on the course of infection.
Lindh
M
M
Department of Clinical Virology, Göteborg University, Sweden. magnus.lindh@microbio.gu.se
Gonzalez
J E
JE
Norkrans
G
G
Horal
P
P
D005838:Genotype; D006515:Hepatitis B virus / Q000145:classification* / Q000235:genetics*; D006801:Humans; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D014773:Virology / Q000379:methods