Epidemiological characteristics of Hepatitis B virus in population in Xianning of Hubei province were investigated by ELISA, conventional PCR detection as well as specific genotyping system by PCR. Blood specimens were detected by ELISA and conventional PCR, the results indicated that 177(11.1%) of 1596 were HBV positive. Comparing with ELISA, conventional PCR is a sensitive, specific method for HBV DNA detection in serum. Positive serum specimens of 53 were identified and differentiated by specific genotyping system by PCR, the results demonstrated that HBV B genotype was found only in population in Xianning and that genotyping system based on PCR using type-specific primers could effectively differentiate HBV genotypes, indicating that this assay system is considered to be a useful tool for the molecular diagnosis of HBV and for large-scale survey of HBV.