The Slide enzyme linked immunosorbent assay (SELISA) was standardized for detection of antibodies specific to Trypanosoma evansi and subsequently used for the screening of naturally infected bovine sera. A novel SELISA, a modification of the standard ELISA technique was used for the detection of antibodies against Trypanosoma evansi in bovines using positive and negative control sera. The test is based on immunostaining of the fixed whole Trypanosoma evansi organisms on microscopic glass slide, incubation with sera, antibovine IgG-HRPO conjugate and substrate Diaminobenzidine tetrahydrochloride (DAB). Finally the reaction was read under oil immersion of microscope. A total of 702 sera samples from bovines in Rayalaseema region of Andhra Pradesh were examined by SELISA and 192 were found positive for Trypanosoma evansi antibodies.