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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1981 Oct;14(4):396–403. doi: 10.1128/jcm.14.4.396-403.1981

BrucELISA: an enzyme-antibody immunoassay for detection of Brucella abortus antibodies in milk: correlation with the Brucella ring test and with shedding of viable organisms.

D K Boraker, W R Stinebring, J R Kunkel
PMCID: PMC271991  PMID: 6793622

Abstract

An indirect enzyme-antibody immunosorbent assay (BrucELISA) is described for the detection of antibody to Brucella abortus in cow's milk. Three series of milk samples were obtained from an adult-vaccinated dairy herd infected with B. abortus. The BrucELISA system was used as a screening test for individual milks diluted 1:200 (BE 200 test), for undiluted bulk milks, and to determine antibody titer (BrucELISA titration assay). The BrucELISA results correlated highly with positive Brucella ring test reactions and culture positivity, eliminated false-positive Brucella ring test reactions, detected antibody in some samples which were Brucella ring test negative, and distinguished between vaccinated and infected animals. BrucELISA titration assay titers of greater than 1:800 were correlated with shedding, or were prognostic for animals which eventually became shedders. Binding of the enzyme-antibody conjugate to bovine immunoglobulin in the absence of rabbit anti-bovine immunoglobulin occurred with culture-positive or -negative milks showing titers of greater than 1:1,600 (the beta effect); the effect was also of predictive value in identifying eventual shedders. The BrucELISA system is a sensitive, specific, and inexpensive method for screening large numbers of individual or bulk milk samples for the presence of antibody to B. abortus.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barta O., Barta V., Miniats O. P., Ingram D. G. Complement and conglutinin in the serum of germ-free and conventional piglets. J Immunol. 1970 Aug;105(2):350–354. [PubMed] [Google Scholar]
  2. Berman D. T., Wilson B. L., Moreno E., Angus R. D., Jones L. M. Characterization of Brucella abortus soluble antigen employed in immunoassay. J Clin Microbiol. 1980 Apr;11(4):355–362. doi: 10.1128/jcm.11.4.355-362.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Byrd J. W., Heck F. C., Hidalgo R. J. Evaluation of the enzyme-linked immunosorbent assay for detecting Brucella abortus antibodies. Am J Vet Res. 1979 Jun;40(6):896–898. [PubMed] [Google Scholar]
  4. CAMERON H. S., KENDRICK J. W. Differentiating postvaccination reactions in brucellosis from virulent infection. J Am Vet Med Assoc. 1957 Jan 15;130(2):90–91. [PubMed] [Google Scholar]
  5. Carlsson H. E., Hurvell B., Lindberg A. A. Enzyme-linked immunosorbent assay (ELISA) for titration of antibodies against Brucella abortus and Yersinia enterocolitica. Acta Pathol Microbiol Scand C. 1976 Jun;84(3):168–176. doi: 10.1111/j.1699-0463.1976.tb00016.x. [DOI] [PubMed] [Google Scholar]
  6. Heck F. C., Williams J. D., Pruett J. Interpretation of spectrophotometric absorbance values to define results of enzyme-linked immunosorbent assays. J Clin Microbiol. 1980 Apr;11(4):398–401. doi: 10.1128/jcm.11.4.398-401.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. INGRAM D. G., BARNUM D. A. FLUCTUATIONS IN THE LEVEL OF CONGLUTININ IN BOVINE SERUM. Can Vet J. 1965 Jul;6(7):162–169. [PMC free article] [PubMed] [Google Scholar]
  8. Lachmann P. J. Conglutinin and immunoconglutinins. Adv Immunol. 1967;6:479–527. doi: 10.1016/s0065-2776(08)60527-1. [DOI] [PubMed] [Google Scholar]
  9. Lamb V. L., Jones L. M., Schurig G. G., Berman D. T. Enzyme-linked immunosorbent assay for bovine immunoglobulin subclass-specific response to Brucella abortus lipopolysaccharides. Infect Immun. 1979 Oct;26(1):240–247. doi: 10.1128/iai.26.1.240-247.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Magee J. T. An enzyme-labelled immunosorbent assay for Brucella abortus antibodies. J Med Microbiol. 1980 Feb;13(1):167–172. doi: 10.1099/00222615-13-1-167. [DOI] [PubMed] [Google Scholar]
  11. Mittal K. R., Jaiswal T. N. Conglutinating activity in apparently healthy human and animal sera. Immunology. 1973 Aug;25(2):237–240. [PMC free article] [PubMed] [Google Scholar]
  12. O'Beirne A. J., Cooper H. R. Heterogeneous enzyme immunoassay. J Histochem Cytochem. 1979 Aug;27(8):1148–1162. doi: 10.1177/27.8.383822. [DOI] [PubMed] [Google Scholar]
  13. O'Sullivan M. J., Bridges J. W., Marks V. Enzyme immunoassay: a review. Ann Clin Biochem. 1979 Sep;16(5):221–240. doi: 10.1177/000456327901600162. [DOI] [PubMed] [Google Scholar]
  14. ROEPKE M. H., PATERSON K. G., DRIVER F. C., CLAUSEN L. B., OLSON L., WENTWORTH J. E. The Brucella abortus ring test. Am J Vet Res. 1950 Apr;11(39):199–205. [PubMed] [Google Scholar]
  15. Roepke M. H., Patterson J. M., Deyoe B. L. Brucella ring test sensitivity of individual and pooled bovine milks with various preservatives. Am J Vet Res. 1974 Jan;35(1):115–118. [PubMed] [Google Scholar]
  16. Roepke M. H., Stiles F. C., Jr Potential efficiency of milk ring test for detection of brucellosis. Am J Vet Res. 1970 Dec;31(12):2145–2149. [PubMed] [Google Scholar]
  17. Ruppanner R., Meyer M. E., Willeberg P., Behymer D. E. Comparison of the enzyme-linked immunosorbent assay with other tests for brucellosis, using sera from experimentally infected heifers. Am J Vet Res. 1980 Aug;41(8):1329–1332. [PubMed] [Google Scholar]
  18. Saunders G. C., Clinard E. H., Bartlett M. L., Sanders W. M. Application of the indirect enzyme-labeled antibody microtest to the detection and surveillance of animal diseases. J Infect Dis. 1977 Oct;136 (Suppl):S258–S266. doi: 10.1093/infdis/136.supplement_2.s258. [DOI] [PubMed] [Google Scholar]
  19. Schuurs A. H., Van Weemen B. K. Enzyme-immunoassay. Clin Chim Acta. 1977 Nov 15;81(1):1–40. doi: 10.1016/0009-8981(77)90410-7. [DOI] [PubMed] [Google Scholar]
  20. Stemshorn B. W., Nielsen K. H., Samagh B. S., Forbes L. B., Ingram D. G. Evaluation of an enzyme-labeled antiglobulin test for anti-Brucella immunoglobulin G among 3 cattle populations. Am J Vet Res. 1980 Nov;41(11):1779–1784. [PubMed] [Google Scholar]
  21. Thoen C. O., Armbrust A. L., Eacret W. G., Harrington R., Jr, Brown G. M. Use of an enzyme immunoassay test for characterizing the A and M antigens of Brucella. J Clin Microbiol. 1979 Apr;9(4):485–487. doi: 10.1128/jcm.9.4.485-487.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thoen C. O., Pietz D. E., Armbrust A. L., Harrington R., Jr Enzyme immunoassay for detecting Brucella antibodies in cow's milk. J Clin Microbiol. 1979 Aug;10(2):222–225. doi: 10.1128/jcm.10.2.222-225.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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