Development of indirect enzyme-linked immunosorbent assay for diagnosis of canine leptospirosis

Research (Published online: 20-05-2017)

11. Development of indirect enzyme-linked immunosorbent assay for diagnosis of canine leptospirosis

A. Sathiyamoorthy, G. Selvaraju, K. M. Palanivel and P. Srinivasan

Veterinary World, 10(5): 530-535

ABSTRACT

Aim: This study was taken up to develop an indirect enzyme-linked immunosorbent assay (i-ELISA) for screening antibodies against Leptospira spp. in canines.

Materials and Methods: An i-ELISA was developed using outer membrane protein extracted from Leptospira interrogans serovar canicola used for coating the well with concentration of 0.5 μg/μl. A total of 250 serum samples from clinically affected and apparently healthy dogs were collected along with relevant epidemiological data at Teaching Veterinary Clinical Complex, Veterinary College and Research Institute, Namakkal, and subjected to i-ELISA.

Results: Out of 250 sera samples, 140 (56.00%) were found to be positive by i-ELISA. All the sera samples were subjected to microagglutination test (MAT) with panel of 12 different serovars. A total of 71 (28.40%) sera samples were positivity to MAT excluding the sera samples positive to L. interrogans serovars canicola and icterohaemorrhagiae in vaccinated dogs. Sensitivity and specificity of i-ELISA were higher in compared with MAT was 91.54% and 58.10%, respectively.

Conclusion: An indirect ELISA developed for the detection of canine antileptospiral antibodies proved to be highly sensitive, rapid and easy to perform and overcome the drawbacks of MAT.

Keywords: canine leptospirosis, indirect enzyme-linked immunosorbent assay, outer membrane protein, Leptospira canicola, Triton X-114 extraction.