Abstract: Objective The study was conducted to investigate the conditions of hepatitis B virus (HBV) infection in Huzhou city, Zhejiang province and its epidemiological characteristics. Methods A total of 1666 randomly selected healthy subjects from the rural mountain areas, plains and regions of rivers and lakes, and towns in Huzhou city participated in the study. Their serum hepatitis B virus surface antigen (HBsAg), hepatitis B virus surface antibody , hepatitis B virus core antibody, hepatitis B virus e antigen. And hepatitis B virus e antibody levels were measured using the enzyme-linked immunosorbent assay. Results The standardized prevalence rate of HBV infection in natural population in Huzhou city was 36.47%, The lowest prevalence rate of hepatitis B was found in the10-year-old group of children, being 4.93%; the HBsAg carrier rate was 4.14%, whereas this rate was 0 in 10-year-olds. The HBV prevalence rate was lower than the national and provincial rates. Conclusion Huzhou has become a moderately prevalent area from a previously highly prevalent one due to the hepatitis B prevalence rate shown in the study. It was revealed that the hepatitis B vaccination had evident effects.