Development of an Individual Rapid Test Based on Enzymatic Immunofiltration Assay for Detection of Anti–Brucella Abortus Antibody in Bovine Sera

Abstract

To detect bovine antibody directed to smooth lipopolysaccharide (LPS), cell lysate (LYS), O-polysaccharide (OPS), and LPS-deprived chromatographic fractions (ChF) of Brucella abortus, 2 bi-antigenic diagnostic models based on the enzymatic rapid immunofiltration assay (ERIFA), ERIFALPS/LYS and ERIFAOPS/ChF, were developed. Their diagnostic performance was compared with complement fixation test (CFT), Rose Bengal test (RBT), indirect in-house and commercial enzyme-linked immunosorbent assays (iELISA and com-ELISA, respectively), based on the smooth LPS antigen, by using a total of 420 cattle sera collected from aborted-unvaccinated, aborted-unvaccinated and culture-positive, healthy-unvaccinated, and healthy-vaccinated cattle. The results demonstrated excellent agreement and no statistical difference between iELISAs and LPS-, LYS-, OPS-based ERIFA models. However, diagnostic performance of CFT, RBT, and ChF-based ERIFA was less significant than that of LPS-, LYS-, and OPS-based ERIFA models, and iELISAs. The results demonstrated a successful adaptation of the multi-antigenic ERIFA model to anti– B. abortus antibody in bovine sera and suggest that the ERIFA model can be considered as an “individual rapid ELISA” due to its similarity with ELISA, individual applicability, and rapidity in determining reactor animals within 5 minutes. In conclusion, the potential of multi-antigenic applications can make the rapid ERIFA model not only an alternative screening method but also a confirmatory test for bovine brucellosis diagnosis.