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Vet Microbiol. 2007 Sep 20;124(1-2):173-7. Epub 2007 Mar 30.

Second generation competitive enzyme immunoassay for detection of bovine antibody to Brucella abortus.

Author information

1
Canadian Food Inspection Agency, Ottawa Laboratory Fallowfield, 3851 Fallowfield Road, Ottawa, Ontario, Canada K2H 8P9. nielsenk@inspection.gc.ca

Abstract

A second generation competitive enzyme immunoassay (CELISA) for detection of bovine antibody to Brucella abortus was developed. This assay was different from previously developed CELISAs in that the detection reagent used was a recombinant combination of the receptor portions of protein A and protein G, labelled with horseradish peroxidase. This eliminates the need for polyclonal anti-mouse-enzyme conjugate reagents for detection thus allowing for true standardization. The assay utilized a monoclonal antibody specific for a common epitope of the O-polysaccharide (OPS) of smooth lipopolysaccharide (SLPS) derived from B. abortus S1119.3 but which did not react with protein A/G. This monoclonal antibody was used to compete with antibody in the bovine test serum. Binding of bovine antibody to the smooth lipopolysaccharide antigen was then measured directly with the protein A/G enzyme conjugate. In this case, development of colour in the reaction was indicative of the presence of bovine antibody. The performance characteristics, sensitivity, specificity and exclusion of B. abortus S19 vaccinated animals, of the assay were very similar to those of the classical CELISA.

PMID:
17467200
DOI:
10.1016/j.vetmic.2007.03.023
[Indexed for MEDLINE]

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