HBV Genotyping

The only commercially available test for genotyping HBV is a line probe assay (Innogenetics, Ghent, Belgium) for mutations in the HBV polymerase gene associated with resistance to lamivudine.18 A 341 base pair (bp) PCR product from the polymerase gene is amplified with primers biotinylated at their 5' ends. The PCR product is denatured and hybridized to probes immobilized on a nitrocellulose strip. The assay uses 38 probes to cover codon positions 528,552, and 555 of the polymerase gene in 19 different probe lines on a nitrocellulose strip. The hybrids are visualized on the strip after addition of strep-tavidin-alkaline phosphatase and colorimetric substrate. The mutations are identified by the colored patterns of PCR product hybridization to the probes.

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