Objective: To establish a simple and practicable method to identify the different genotypes of hepatitis B virus (HBV).
Methods: Based on the alignment of 114 complete nucleotide sequence for HBV DNA of different genotypes, the specific sequence of each genotype was found. Six primer sets were designed for each of the six genotypes according to the genotype-specific sequence, and used separately for PCR. The genotype of HBV was identified according to the positive result of PCR. Three primer sets for B, C and D genotypes were added into a single tube for PCR reaction, and HBV was genotyped according to the length of the amplified DNA.
Results: There was no difference in the genotyping result of PCR by single or multiplex primers, which was identical to the PCR-RFLP method.
Conclusions: This multiplex PCR method is simple, precise, sensitive, and easy to use.