The transmission of infectious agents such as human immunodeficiency virus (HIV), hepatitis B (HBV), and hepatitis C (HCV) represents the greatest threat to the safety of blood transfusion to the recipient. The association of HIV infection with HBV and/or HCV is common worldwide, due to shared modes of transmission. The prevalence of HIV-HBV co-infection is estimated to be 5-10% in the United States and 20-30% in Asia and parts of sub-Saharan Africa. Other studies report HCV co-infection in 9-25% of HIV-infected patients. The aim of this study was to determine the frequency of markers of co-infection between hepatitis (B,C) and human immunodeficiency virus in blood donations collected at the CHU Kati blood transfusion center. It was a retrospective study conducted from October 1 to December 31, 2018. Data were collected from blood donors aged 18 to 60 years. The Pre-donation medical interview was the first barrier for the selection of subjects at risk. The sample was composed of 92% males and 8% females out of a total of 507 individuals sampled. Biological screening was performed by ELISA (Enzyme Linked Immuno Sorbent Assay). The results revealed a prevalence of 0.86% of HIV-HBV co-infection only in rhesus negative donors. No co-infection between HIV and HCV was found. This study showed that at the Kati University Hospital, only rhesus-negative donors presented cases of HIV-HBV co-infection.