Recombinant LipL32 and LipL41 outer membrane proteins of Leptospira interrogans serovar Canicola were produced, and used as a pooled antigen in enzyme-linked immunosorbent assay (ELISA) to detect leptospiral antibodies in bovine sera samples. The optimum concentration of the pooled antigen was found to be 50ng of each antigen per well by using known positive and negative cattle sera. Using a total of 500 bovine sera samples the sensitivity, specificity and accuracy of pooled antigen based ELISA as compared to microscopic agglutination test (MAT) were 100%, 88.1% and 91.6%, respectively. The results suggested that antigen in ELISA could be preferred for detection of all those cases, which might have remained undiagnosed by performing MAT.

A total of 281 serum samples collected randomly from goats showing the signs of fever, abortion,repeat breeding and still births as well as from apparently healthy ones were subjected to LAT and MAT based on rLipL32 and rLipL41 antigens. A total of 16 (5.69%) samples were found positive to MAT, whereas rLipL32-LAT and rLipL41-LAT detected 35 (12.45%) and 23 (8.18%) samples as positive, respectively. The sensitivity and specificity of rLipL32-LAT was 87.50% and 92.83%, respectively,while rLipL41-LAT yielded 75.00% and 97.35% sensitivity and specificity, respectively. LAT based on rLipL32 and rLipL41antigens could further be evaluated on a larger number of samples to ensure its utility as a screening test for the sero-epidemiological studies.