Lateral flow assay for rapid serodiagnosis of bovine leptospirosis

Background:Leptospirosis is considered to be an economically important disease in bovine. The disease burden is not appropriately monitored due to cumbersome serological tests that could be performed only in established laboratories. This warrants the development of a field level rapid diagnostic test. Aims:The study aimed to develop a lateral flow assay (LFA)-based pen-side diagnostic test to detect antibodies to Leptospira. Methods:LFA strip was prepared with the heat extracted antigen from L. interrogans serovar Pomona. To assess the performance of the developed LFA, a total of 300 bovine serum samples with their clinical histories were used and the initial screening for Leptospira antibodies was performed by the standard microscopic agglutination test (MAT). The sensitivity, specificity, and agreement (kappa value) were calculated between developed LFA and MAT. The stability of LFA was evaluated on days 30, 60, 90, and 120. Results:Out of 300 samples tested, 225 were positive, and 75 were negative on MAT and 208 were positive, and 92 were negative on LFA. The developed LFA had a sensitivity of 90.7% and a specificity of 94.7%. The results of the assay were substantially in agreement with MAT, with a kappa value of 0.79. The LFA strips were stable for 120 days at 4°C. Conclusion:A Lateral flow assay-based rapid pen-side test was developed and its utility to diagnose bovine leptospirosis was evaluated.Key Words: Bovine, Immunochromatography, Lateral flow assay, Leptospirosis