The use of divalent cation chelating agents (EDTA/EGTA) to reduce non-specific serum protein interaction in enzyme immunoassay

An indirect enzyme immunoassay (ELISA) for detection of bovine antibody to Brucella abortus was modified by the addition of divalent chelating agents to the serum diluent. This addition resulted in an increase in specificity from 96.0% in the regular assay to 99.4% in the modified procedure. Of the 15,715 sera initially tested by the indirect ELISA, 691 that had given positive reactions were selected for retesting in the indirect ELISA with EDTA/EGTA added. The buffered plate antigen test (BPAT) correctly identified 98.6% of the samples as negative. The addition of chelating agents did not alter the sensitivity of the indirect ELISA, which correctly classified 609 sera from animals from which B. abortus had been isolated as positive. The sensitivity of the BPAT was 97.8%.