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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Dec;33(12):3198–3200. doi: 10.1128/jcm.33.12.3198-3200.1995

Evaluation of PCR and indirect enzyme-linked immunosorbent assay on milk samples for diagnosis of brucellosis in dairy cattle.

C Romero 1, M Pardo 1, M J Grillo 1, R Diaz 1, J M Blasco 1, I Lopez-Goñi 1
PMCID: PMC228673  PMID: 8586702

Abstract

A study was performed to evaluate the previously described PCR (C. Romero, C. Gamazo, M. Pardo, and I. López-Goñi, J. Clin. Microbiol. 33:615-617, 1995) for the diagnosis of brucellosis in dairy cattle. Milk samples from 56 Brucella milk culture-positive cattle and from 37 cattle from Brucella-free herds were examined for Brucella DNA by PCR and for specific antibodies by an indirect enzyme-linked immunosorbent assay (ELISA). The specificities of both tests were 100% when testing the milk samples from Brucella-free cattle. The milk samples from 49 infected cattle were positive by PCR (87.5% sensitivity), and 55 were positive by ELISA (98.2% sensitivity). A PCR-positive sample was negative by ELISA, and 7 ELISA-positive samples were PCR negative, yielding an observed proportion of agreement of 0.91 for the two tests. Although the results suggest that ELISA is a better screening test than PCR, the combined sensitivity of the two assays was 100%, and their simultaneous application could be more useful than one test alone for a rapid screening of brucellosis in dairy cattle.

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Selected References

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  1. Baily G. G., Krahn J. B., Drasar B. S., Stoker N. G. Detection of Brucella melitensis and Brucella abortus by DNA amplification. J Trop Med Hyg. 1992 Aug;95(4):271–275. [PubMed] [Google Scholar]
  2. Boraker D. K., Stinebring W. R., Kunkel J. R. 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. J Clin Microbiol. 1981 Oct;14(4):396–403. doi: 10.1128/jcm.14.4.396-403.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown G. M., Ranger C. R., Kelley D. J. Selective media for the isolation of Brucella ovis. Cornell Vet. 1971 Apr;61(2):265–280. [PubMed] [Google Scholar]
  4. Dorsch M., Moreno E., Stackebrandt E. Nucleotide sequence of the 16S rRNA from Brucella abortus. Nucleic Acids Res. 1989 Feb 25;17(4):1765–1765. doi: 10.1093/nar/17.4.1765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Díaz-Aparicio E., Marín C., Alonso-Urmeneta B., Aragón V., Pérez-Ortiz S., Pardo M., Blasco J. M., Díaz R., Moriyón I. Evaluation of serological tests for diagnosis of Brucella melitensis infection of goats. J Clin Microbiol. 1994 May;32(5):1159–1165. doi: 10.1128/jcm.32.5.1159-1165.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Farrell I. D. The development of a new selective medium for the isolation of Brucella abortus from contaminated sources. Res Vet Sci. 1974 May;16(3):280–286. [PubMed] [Google Scholar]
  7. Fekete A., Bantle J. A., Halling S. M. Detection of Brucella by polymerase chain reaction in bovine fetal and maternal tissues. J Vet Diagn Invest. 1992 Jan;4(1):79–83. doi: 10.1177/104063879200400118. [DOI] [PubMed] [Google Scholar]
  8. Fekete A., Bantle J. A., Halling S. M., Sanborn M. R. Preliminary development of a diagnostic test for Brucella using polymerase chain reaction. J Appl Bacteriol. 1990 Aug;69(2):216–227. doi: 10.1111/j.1365-2672.1990.tb01512.x. [DOI] [PubMed] [Google Scholar]
  9. Herman L., De Ridder H. Identification of Brucella spp. by using the polymerase chain reaction. Appl Environ Microbiol. 1992 Jun;58(6):2099–2101. doi: 10.1128/aem.58.6.2099-2101.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Huber J. D., Nicoletti P. Comparison of the results of card, rivanol, complement-fixation, and milk ring tests with the isolation rate of Brucella abortus from cattle. Am J Vet Res. 1986 Jul;47(7):1529–1531. [PubMed] [Google Scholar]
  11. Kerkhofs P., Botton Y., Thiange P., Dekeyser P., Limet J. N. Diagnosis of bovine brucellosis by enzyme immunoassay of milk. Vet Microbiol. 1990 Jul;24(1):73–80. doi: 10.1016/0378-1135(90)90052-w. [DOI] [PubMed] [Google Scholar]
  12. Nakane P. K., Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem. 1974 Dec;22(12):1084–1091. doi: 10.1177/22.12.1084. [DOI] [PubMed] [Google Scholar]
  13. Romero C., Gamazo C., Pardo M., López-Goñi I. Specific detection of Brucella DNA by PCR. J Clin Microbiol. 1995 Mar;33(3):615–617. doi: 10.1128/jcm.33.3.615-617.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Thoen C. O., Bruner J. A., Luchsinger D. W., Pietz D. E. Detection of brucella antibodies of different immunoglobulin classes in cow milk by enzyme-linked immunosorbent assay. Am J Vet Res. 1983 Feb;44(2):306–308. [PubMed] [Google Scholar]

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