Development of OMP based indirect ELISA to gauge the antibody titers in bovines against Pasteurella multocida

Document Type: Full paper (Original article)

Authors

1 MVSc in Veterinary Microbiology, Department of Veterinary Microbiology, Dr. G. C. Negi College of Veterinary & Animal Sciences, CSK Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, 176062, India

2 Department of Veterinary Microbiology, Dr. G. C. Negi College of Veterinary & Animal Sciences, CSK Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, 176062, India

3 Department of Veterinary Physiology & Biochemistry, Dr. G. C. Negi College of Veterinary & Animal Sciences, CSK Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, 176062, India

Abstract

Pasteurella multocida (P. multocida) is an important pathogen of various domestic animals. The outer membrane proteins (OMPs) play a major role in pathogenesis and immunogenicity of P. multocida. The aim of the study was to develop indirect enzyme linked immuno sorbant assay (ELISA) based on OMPs to ascertain the antibody titers in animals post-infection or to gauge the potency of vaccine. The OMPs were extracted and purified from P. multocida P:52 (vaccine strain) and P. multocida B:2 isolated from natural outbreak of Haemorrhagic septicaemia (HS) and analyzed on SDS PAGE and through western blot. The OMPs profile of the vaccine strain and the isolate from the natural outbreak of HS were found to be similar. Optimization of various components viz. coating antigens, anti-species conjugate, etc. were carried out against both anti-P. multocida hyper immune and pre immune serum. Validation of OMP based indirect ELISA assay to measure immune response against P. multocida in bovine revealed 91% diagnostic sensitivity (DSN) and about 100% diagnostic specificity (DSP) at 25% cut off. OMP based indirect ELISA was found to be more specific, but less sensitive as compared to WCL based assay.

Keywords