This study aims to determine the prevalence of human immunodeficiency virus (HIV) infections and hepatitis B virus (HBV) and hepatitis C virus (HCV) infections among blood donors at the Blood Donor Center, Mohammed V Military Teaching Hospital between 2010 and 2012. We conducted a retrospective study among military blood donors aged 18-50 years, with a male predominance (95%). Pre-donation interview is the first selection barrier for individuals at risk. Biological screening was performed by liquid enzyme immunoassay technique using antibodies and/or antigen. Fourth generation combined HCV and HIV antigen/antibody ELISA (enzyme-linked immunosorbent assay) test was used. The Blood Donor Center and the laboratory of virology used the same technique performed in duplicate to confirm results. Out of 25661 tested samples, the prevalence rate of HBV infections was 3.97 % (n = 102), the prevalence rate of HCV infections was 2.45 % (n = 63) and the prevalence rate of HIV infections was 0.15 % (n = 4). A single case with HBV and HCV virus co-infection (0.039 %) was registered, no association between HIV-HBV, HIV-HCV or HBV, HCV and HIV infections was recorded. The low seroprevalence rates of viral markers recorded in our study show improvement in preventive measures for donor selection and screening tests. The registered prevalence encourages the use of combined reagent, which is the only alternative to molecular biology in developing countries.