Development of Qualitative and Quantitative ELISA Models for Bovine Brucellosis Diagnosis

Abstract

In this study, a qualitative and a quantitative ELISA models (iELISA and qELISA, respectively) have been developed for screening and quantifying IgG isotype anti-Brucella abortus LPS antibody. For developing high sensitive and specific ELISA models, the nature and concentration of blocking-diluting reagents have been found very critical since milk components such as casein and particularly β-lactoglobulin, but not α-lactalbumin, strongly inhibited the anti-LPS antibody detection by an unknown mechanism. The iELISA as well as qELISA models have been developed and validated by OIE reference sera and well-known field sera evaluated with RBT, CRT and cELISA. The results demonstrated that both ELISA models, iELISA in large-scale screening and qELISA in standard determination of anti-LPS antibody concentration, developed in this study can be included as valuable and effective immunodiagnostic tools in brucellosis monitoring-eradication and vaccination surveillance programs in endemic countries.