Development and Validation of a Competitive ELISA Kit for the Serological Diagnosis of Ovine, Caprine and Bovine Brucellosis

Abstract

A competitive ELISA (Brucella-Ab c-ELISA) was standardized and validated for the detection of Brucella antibodies in cattle, sheep and goat sera using a monoclonal antibody (MAb 4B5A) produced against Brucella melitensis biotype 2. The specificity and sensitivity of the assay were 100% to a 67.5% cut-off point (B/Bo%). When compared with an indirect ELISA, the Brucella-Ab c-ELISA did not demonstrate cross-reactions when testing positive sera for antibodies to some Enterobacteriaceae. A comparison was made between the Brucella-Ab c-ELISA and the complement fixation and Rose Bengal tests. Results demonstrated that the Brucella-Ab c-ELISA is a valuable tool for the serological diagnosis of bovine and ovine/caprine brucellosis.

Field evaluation of fluorescence polarization assay, and comparison with competitive ELISA for the detection of antibodies against Brucella melitensis in sheep in Sicily, Italy

“…The very high accuracy of c-ELISA that we found in this study is in agreement with previous findings (Portanti et al, 2006). Indeed, although FPA was shown as an accurate and rapid test, c-ELISA appears to be the superior in terms of Se, which is a critical factor in the eradication of B. melitensis in Southern Italy.…”

“…Accuracy of FPA was, then, compared with that of c-ELISA, which is a currently available, rapid test, which was previously proposed for improving accuracy and reducing costs of antibody testing for small ruminant brucellosis in Italy (Portanti et al, 2006).…”

Herd prevalence of bovine brucellosis and analysis of risk factors in cattle in urban and peri-urban areas of the Kampala economic zone, Uganda

“…screening tests are known to be caused by unrelated Enterobacteriaceae [26-35] and CELISA can eliminate such reactions [33]. B. abortus vaccination strain 19 also gives rise to an antibody response similar to that resulting from natural infection [33] but CELISA can eliminate this false-positive reaction only by approximately 50% [36]. In our study, 9/177 (5.1%) of farms held vaccinated cattle and the CELISA positive reactions of vaccinated cows in two farms might be due to the false-positive reactions with B. abortus vaccination strain S19.…”

Rapid immunofiltration assay based on colloidal gold–protein G conjugate as an alternative screening test for bovine and ovine brucellosis

“…1 Representative results of the NERIFA using positive and negative serum samples present study was performed with undiluted well-identified bovine and ovine sera. The NERIFA model was first optimized by using a mouse monoclonal anti-Brucella LPS antibody (clone 4B5A, Portanti et al 2006) and a few number of Brucella antibody positive and negative sera. As seen in Fig.…”