The rapid diagnostic tests used in most resource-limited countries offer little specificity in the differentiation of HIV-1, HIV-2 and HIV-1 + 2. World Health Organization (WHO) does periodically evaluate rapid tests in use in the South Countries. Despite the pre-qualification from WHO, it is necessary at local level to conduct comparative studies between the available tests. To do this in Mali, we conducted a cross-sectional study at the University Teaching Hospital Gabriel Touré and the Research and Training Center (SEREFO) of Bamako, on 172 samples from retrospective and prospective studies. The goal of this project was to compare the following rapid tests: “HIV TRI-DOT” and OnSite HIV1/2 Ab Plus Combo Rapid Test for screening HIV-1 and HIV-2 to “ImmunoComb II BiSpot” (gold standard). The sensibility and specificity of “HIV TRI-DOT” for HIV-1 detection were 100 % each [IC 95: 87, 13-100 and 95,31-100]. Its performance for HIV-2 detection was 95.24% [IC 95: 84,21-98,68] (sensibility) and 100% [IC 95: 72,25-100] (specificity). The positive and negative predictive values were respectively 100% [IC 95: 91,24-100] and 83.33% [IC 95: 55.20-95,30]. The sensibility and specificity of OnSite HIV1/2 Ab Plus Combo Rapid Test assay performance were 100% [IC 95: 87,13-100 and 95,31-100]. Overall, OnSite HIV ½ Ab Combo Rapid Test proved to be a good test for warm countries and does not require refrigeration in these settings. Blood and its products can be used to perform the test, unlike “ImmunoComb II BiSpot” and “HIV TRI-DOT” which must be kept cold and cannot be done with whole blood.