A universal hepatitis B vaccination programme has been conducted in Long An county since 1986. To investigate the epidemiological changes in hepatitis B virus (HBV) infection we conducted a serosurvey there in 2005. A total of 4686 subjects were enrolled and vaccination history and blood samples collected. HBV infective markers were determined by radioimmunoassay. The results were compared with the data of 1985. Our results show that the overall HBsAg prevalence was 7·5%, less than half of the prevalence reported in 1985. HBsAg and anti-HBc antibody prevalence in people born after 1985 decreased markedly. The gender difference in HBsAg prevalence was abolished in subjects aged <20 years. The administration of a first dose of vaccine within 24 h could reduce the HBsAg prevalence by half. In conclusion, the marked epidemiological changes in HBV prevalence found in this serosurvey indicate that the implementation of HBV vaccination was highly successful.