Improved multiplex-PCR to identify hepatitis B virus genotypes A–F and subgenotypes B1, B2, C1 and C2

BackgroundThere are eight genotypes (A–H) and numerous subgenotypes of hepatitis B virus (HBV). The genotype has been shown to affect the course of HBV infection.ObjectivesTo develop an efficient genotyping and subgenotyping method for large-scale epidemiological surveys of HBV infection in countries with high prevalence of HBV B and C such as China.Study designWe designed genotype and subgenotype-specific primer pairs, and adjusted PCR conditions for a multiplex-PCR using common Taq polymerase to identify HBV genotypes A–F in one reaction and for the main subgenotypes B1/B2 and C1/C2 in another reaction.ResultsWe have developed a multiplex-PCR system, which specifically amplifies DNA of HBV genotypes and the corresponding main subgenotypes B and C from the sera of HBV patients. Our patients were infected with HBV of subgenotypes B2 (n = 18), C1 (n = 2) and C2 (n = 48). Eleven patients were doubly infected and three showed a triple infection with HBV A, B and C.ConclusionsThe low-cost multiplex-PCR for identification of HBV genotypes A–F and main subgenotypes of HBV B and C, is rapid, reliable and sufficient for large-scale epidemiological surveys and clinical studies.