[Application of multiplex PCR in genotyping of hepatitis B virus prevailing in Guangdong Province of China]

Abstract

Objective: To establish a convenient method for the genotyping of hepatitis B virus (HBV) using multiplex PCR.

Method: Based on the alignment of 114 complete nucleotide sequences of HBV DNA belonging to different genotypes, acquired from the GenBank, genotype-specific sequences were identified according to which 6 pairs of primers were designed corresponding to each genotype. Subsequent genotyping of HBV was performed using these primers that were added, either alone or in conjunction with others, into a multiplex PCR reaction tube, and HBV genotype was determined according to the length of amplified DNA.

Result: The genotyping result of multiplex PCR was consistent with that produced by PCR- restriction fragment length polymorphism as established by Lindh. We found in this study that among the HBV carriers in the vicinities Guangzhou of City, about 45% belonged to B genotype, 38.75% to C genotype and 16.75% to D genotype.

Conclusion: This multiplex PCR method is simple, convenient and more differential.