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Introduction: Sub-Saharan Africa struggles continuously with insufficient resources and inadequate infrastructure that hinder the establishment of a safer blood supply despite improvements in transfusion safety over recent decades. This study aimed to evaluate the impact of the chemiluminescence technique in combination with immunoenzymatic and immunochromatographic tests for viral marker screening of hepatitis B (HBV), hepatitis C (HCV) and human immunodeficiency virus (HIV) in donated blood in a country of sub-Saharan Africa. Method: This study was conducted in a population of 113,406 blood donors at the National Centre of Blood Transfusion in Senegal. The data were obtained from the ‘INLOG’ software and donor registers. Statistical analyses used Excel 2010 and Epi Info v6. Screening for HBsAg viral markers, anti-HCV Ab, HIV p24 Ag, anti-HIV1 and anti-HIV2 antibodies were first carried out using the chemiluminescence technique. Blood donations screened positive for HBV or HCV were retested in a second chemiluminescence equipment. HIV-positive donations and their controls were subjected to solid phase immunochromatographic and indirect enzyme immunoassay techniques. Results: The prevalence among donors of HBV was 8.39 %, 0.56 % for HCV and 0.18 % for HIV. Of the donors tested positive for HIV in screenings and in doubled-controls, only 61.54 % were confirmed by the alternative tests; 34.02 % were negative and 4.44 % discordant between the three techniques. Conclusion: This study shows the importance of introducing the chemiluminescence technique in association with serological screening of transfusion-transmitted viruses to improve blood supply safety in low-income countries.