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Veterinary Research Communications

, Volume 18, Issue 6, pp 433–437 | Cite as

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

  • K. Nielsen
  • L. Kelly
  • D. Gall
  • P. Smith
  • J. Bosse
  • P. Nicoletti
  • W. Kelly
Immunology

Abstract

An indirect enzyme immunoassay (ELISA) for detection of bovine antibody toBrucella 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 whichB. abortus had been isolated as positive. The sensitivity of the BPAT was 97.8%.

Keywords

Brucella abortus cattle chelating agents EDTA EGTA ELISA immunoassay specificity 

Abbreviations

ABTS

2,2′-azinobis(3-ethylbenzthiazoline sulphonic acid)

BPAT

buffered plate antigen test

EDTA

ethylenediaminetetraacetic acid disodium salt

EGTA

ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

ELISA

enzyme-linked immunosorbent assay

IgG1

immuno- globulin G1

IgM

immunoglobulin M

Tris

tris(hydroxymethyl)aminomethane

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References

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Copyright information

© Kluwer Academic Publishers bv 1994

Authors and Affiliations

  • K. Nielsen
    • 1
  • L. Kelly
    • 1
  • D. Gall
    • 1
  • P. Smith
    • 1
  • J. Bosse
    • 1
  • P. Nicoletti
    • 2
  • W. Kelly
    • 1
  1. 1.Animal Diseases Research InstituteAgriculture CanadaNepeanCanada
  2. 2.Department of Infectious Diseases, College of Veterinary MedicineUniversity of FloridaGainesvilleUSA

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