National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
The present study aims at evaluating the efficacy of two rapid tools, IgM dot ELISA and Latex Agglutination Test employing recombinant LipL32 as antigen in diagnosing acute leptospirosis in dogs in Kerala, a disease endemic area. One hundred and five serum samples were collected from dogs presented at Veterinary Hospitals attached to Kerala Veterinary and Animal Sciences University during the period from September 2014 to May 2016. Microscopic Agglutination test was carried out using the reference strains maintained in the Dept. of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy. In MAT, samples with titre ≥ 1:800 were considered as positive. For recombinant LipL32 protein production, the lipl32 gene of Leptospira interrogans was ligated with prokaryotic expression vector pPro.EX.HtC and transformed into competent E. coli DH5α cells. The clones were induced with Isopropyl β-D-1-thiogalactopyranoside (IPTG) and the protein was purified using Nickel affinity chromatography and used as antigen in IgM dot ELISA, plate ELISA and Latex Agglutination Test (LAT). All the 105 samples were subjected to the three tests. The relative sensitivity and specificity of IgM dot ELISA, plate ELISA and LAT were evaluated in comparison with MAT. Among the 105 samples, 56 were found to have a titre of ≥1:800 in MAT. In IgM dot ELISA, 55 samples were found to be positive and one was negative. Latex Agglutination Test detected 58 samples as positive. It was found to be a less specific (91.84 %) test than IgM dot ELISA in detecting acute leptospirosis although it was found to be sensitive (96.43 %). IgM dot ELISA was proved to be sensitive (96.43 %) and specific (97.96 %) and the results were more satisfactory than IgM plate ELISA in detecting acute cases of leptospirosis which is very relevant in an endemic area like Kerala.
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