[en] Visceral leishmaniasis (V L) is endemic in the Sudan where it poses a major health problem in many parts of the country. This study was designed to evaluate parasitological, immunological and DNA based detection methods for diagnosis of V L in Sudan. Direct agglutination test was performed in sera from V L suspects (n,513) using the trypsin treated antigen DAT.T of sensitivity 89%. Moreover, DAT.T detected V L in 43% of unconfirmed V L cases (P<0.001). The sensitivity of in vitro culture was obtained as 40% in comparison to 98% for DAT.T. The sensitivity of 2ME-DAT.R was 95.8% in comparison to 96.3% for DAT.T (P>0.05). The detection rate of 2ME-DAT.R and DAT.T, in unconfirmed (n,67), was 46.3% and 40.3%, respectively (P>0.05). The specificity of both DAT antigens, in non-endemic controls (n,200), was 100%. The specificity of both DAT.R and DAT.T, in sera of patient with other diseases (n, 178), was 98.9% and 98.3%, respectively. The sensitivity of 2ME-DAT.R, using specimens of serum, saliva and urine was 100%, 21.7% and 0%, respectively. DAT.R was positive in saliva and sera of 2 unconfirmed V L cases. The sensitivity of slide formol gel test (FGT) was 84.3% in comparison to 89% for DAT.T (P<0.05). The detection rate for FGT and DAT.T, was 51.3% and 43.3%, respectively (P<0.05). The specificity of FGT, in relation to non-endemic controls (n, 200), was 100%. Nevertheless, FGT specificity with other diseases, was 94%. FGT was positive in 23 of PKDL suspects (n, 35). FGT showed negative reactions in 39%, and lower reactions in 29%, of V L confirmed cases (n, 69) immediately after treatment. The sensitivity of latex agglutination test (LAT), using L.donovani soluble antigen, was 79% in comparison to 98.7% for DAT.T (P<0.001). The detection rate in unconfirmed cases (n, 26) for LAT and DAT.T was 39% and 46%, respectively (p<0.001). The specificity of LAT non-endemic controls (n, 27), was 96%. LAT cross-react with a serum of onchocerciasis (n, 5). The sensitivity of LAT L. donovani lyophilized antigen, was 70% in comparison to 96.6% for DAT (p<0.001). The detection rate for LAT and DAT.T, in unconfirmed cases (n, 23), was 78% and 30%, respectively. The specificity of LAT, in non-endemic controls (n, 30), was 70% in comparison to 100% for DAT.T. The LAT L. donovani lyophilized antigen was positive in 85% of PKDL suspects, in comparison to 61% by DAT. Heat inactivation of sera was found to increase the LAT specificity from 70% to 100%, the LAT sensitivity from 79% to 96%. The sensitivity of LAT L.infantum soluble antigen was 29%, which was less than DAT.T (100%) (P<0.001). The detection rate of LAT, in unconfirmed cases (n, 23), was zero in comparison to 30% by DAT (P<0.05). The specificity of LAT in controls (n, 30) was 100%. All PKDL suspects (n, 13) were negative in LAT while in DAT was positive in eight of them. The sensitivity of counter immuno electrophoresis (CIE) and DAT.T, was obtained as 64% and 100%, respectively (P>0.05). The detection rate of CIE and DAT.T, was 33% in comparison to 22% by DAT.T. The specificity of CIE and DAT.T was 100%. The sensitivity of cellulose acetate perciptin (CAP) was 33%, while that of DAT.T was 100% (P<0.05). The detection rate for CAP and DAT was 11% and 22%, respectively. The specificity of CAP controls (n, 11) was 100%. The sensitivity of Pcr using blood and bone marrow specimens in comparison to DAT.R, was 33%, 100% and 89%, respectively. The detection rate for Pcr using blood and bone marrow, was 45.5% and 91%, respectively in comparison to 91% for DAT.R. The specificity of Pcr blood and DAT.R was 100%. DAT.R is more sensitive than Pcr using blood (P<0.05). But, statistically there is no significant difference between DAT.R and Pcr using bone marrow (P>0.05)