Objective: To understand hepatitis B virus(HBV) infection in high school students in Guangxi, and to provide scientific foundation for hepatitis B prevention and control strategies. Methods: A selfdesigned epidemiological questionnaire was used among 2 632 participants. Information regarding general demographic characteristics, lifestyle habits, HBV infection history, knowledge of viral hepatitis prevention and control, as well as hepatitis B vaccination. Univariate and multivariate logistics regression analysis was used to analyze associated factors for hepatitis B infection. Results: A total of 192 out of 2 632 participants were found HBsAg positive and 2 440 were HBsAg negative. Multivariate analysis showed that family liver disease history and hairdressing injury positively associated with hepatitis B virus infection risk (OR=3.62, 95% CI=2.28-5.73;OR=3.06, 95%CI=1.94-4.83), and hepatitis B vaccination was negatively associated with hepatitis B virus infection risk (OR=0.08, 95%CI=0.05-0.11). Conclusion Family history of liver disease, hairdressing injury experiences, as well as low rate of hepatitis B vaccination may associate with relatively high prevalence of hepatitis B virus infection among high school students in Guangxi.