In developing countries or economies in transition, many people die because of lack of safe blood, even in some urban healthcare facilities. Blood transfusions can save lives and improve health, but millions of patients did not timely access to safe blood, due to no reliable blood donors. The objective of this study was to determine the prevalence of hepatitis B in blood donors from the Sendwe General Provincial Hospital. This is a retrospective cross-sectional descriptive study on serum markers of hepatitis B study taking place over a period of 12 months from 1 January to 31 December 2014. This study showed that the prevalence of hepatitis B was 6.8%, and 78.7% of donors were male. The mean age was 34.1 ± 7.4 years, ranging from 18 to 57 years. Married, family donors, blood type O and Rh positive were the most represented with 81.1%, 68.6%, 58.8% and 99.3% respectively. A significant association was observed between the prevalence of hepatitis B and age, civil status of blood donors (p ≤ 0.001). These results should encourage health authorities to the implementation of effective prevention methods to stop the rapid spread of this infection, which is a serious public health problem.