Evaluation of recombinant LigB antigen-based indirect ELISA and latex agglutination test for the serodiagnosis of bovine leptospirosis in India

Mol Cell Probes. 2014 Aug;28(4):141-6. doi: 10.1016/j.mcp.2014.01.001. Epub 2014 Jan 18.

Abstract

Leptospirosis is a zoonotic disease caused by pathogenic spirochetes of the genus Leptospira, causing febrile infection characterized by multi-organ failure in humans and animals. Leptospiral Ig-like protein B (LigB) is a surface-expressed antigen that mediates host cell invasion or attachment. In this study, N-terminal conserved region of LigB protein (46 kDa) was evaluated for its diagnostic potential to detect anti-leptospiral antibodies in the sera of various animal species. Dot blot analysis revealed immunoreactivity of Leptospira-positive sera of cattle, buffalo, dog, sheep and goat to purified LigB protein. We have analyzed 1126 bovine serum samples, collected from Northern and Eastern part of India, by microscopic agglutination test (MAT) and recombinant LigB (rLigB) based ELISA and latex agglutination test (LAT). The sensitivity of rLigB based ELISA for 554 MAT positive sera was 96.9% and the specificity with 572 MAT negative sera was 91.08% whereas LAT showed sensitivity and specificity of 93.68% and 92.31%, respectively. Kappa values of 0.879 and 0.860 for recombinant antigen based ELISA and LAT indicate excellent agreement with the gold standard serological test, MAT, for the detection of anti-leptospiral antibodies in sera. Further, LAT based on rLigB antigen is a simple and rapid test, suitable for serodiagnosis of leptospirosis under field conditions, owing to its portability and longer shelf life.

Keywords: ELISA; Latex agglutination test; Leptospirosis; LigB; Microscopic agglutination test.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood*
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology
  • Cattle
  • Cattle Diseases / blood
  • Cattle Diseases / diagnosis
  • Cattle Diseases / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods
  • India
  • Latex Fixation Tests / methods
  • Leptospirosis / diagnosis
  • Leptospirosis / immunology*
  • Leptospirosis / veterinary*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Zoonoses / blood
  • Zoonoses / diagnosis
  • Zoonoses / immunology

Substances

  • Antibodies, Bacterial
  • Antigens, Bacterial
  • LigB-m protein, Leptospira interrogans
  • Recombinant Proteins