Hepatitis B virus (HBV) infection is hyperendemic in Cameroon, and healthcare 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. 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. We enrolled 1824 of 1836 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 < 0.001). In conclusion, approximately 1 in 12 HCWs in Cameroon have evidence of HBV infection, yet fewer than 1 in 6 have been fully vaccinated. Our results illustrate the urgent need to scale up systematic HBV screening and targeted vaccination of HCWs in the region.