INTRODUCTION: Hepatitis B virus (HBV) infection is hyperendemic in Cameroon, and health care workers (HCWs) are at high-risk of infection. We aimed to assess prevalence, risk factors and vaccine coverage of HBV infection among HCWs in Cameroon. METHODS: We conducted a cross-sectional study in 16 hospitals across all regions of Cameroon. HCWs were tested for HBV using rapid diagnostic tests (RDT). We collected data on socio-demographics and HBV vaccination status. We estimated prevalence of HBV and used Poisson regression models with robust standard errors to model the prevalence ratios of HBV positivity between covariates. RESULTS: We enrolled 1,824 of 1,836 eligible HCWs (97.5%). The mean age was 34 (SD: 10) years, 65.3% (n=1787) were women, and 11.4% (n=1747) had three or more doses of the HBV vaccine. Overall, we found a HBV prevalence of 8.7% (95% CI: 5.2 – 14.3%). Patient transporters had the highest crude prevalence (14.3%; 95%CI: 5.4–32.9%), whereas medical doctors had the lowest (3.2%; 95%CI: 0.8%−12.1%). The Far North Region had the highest prevalence of HBV (24.0%; 95%CI: 18.3%−30.8%). HBV prevalence decreased with increasing doses of the HBV vaccine (10.3% for no doses vs 3.5% for three or more doses; P