Blood transfusion is a therapeutic act which consists of administering blood or one of its components (red blood cells, platelets and plasma), from one or more healthy subjects to a sick subject. Blood donation is a life-saving gesture that is highly encouraged but a source of pathologies which constitutes a real public health problem. Objective: To study the seroprevalence of viral markers among blood donors at the Gabriel Touré University Hospital in Mali. Method: Cross-sectional study from January 2020 to December 2021. This study was carried out at the CHU Gabriel Touré transfusion service in Mali on the results of blood donors. The donors were selected according to the criteria established by the National Blood Transfusion Center in Mali. Results: the prevalence of HIV, HBV, HCV and Syphilis were respectively 0.27%, 10.67%, 2.28% and 0.76% among all donors. The male gender dominated with a positive prevalence of 10.98% for HBV. The prevalence of HIV and Hepatitis B co-infection was the most represented with 0.27% collected. Conclusion: the prevalence of infectious agents among blood donors remains high in Bamako, Mali. This study suggests greater attention to raising awareness about testing for viral agents like HIV and hepatitis B.