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Journal of Clinical Microbiology, February 2005, p. 721-725, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.721-725.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Indirect Enzyme-Linked Immunosorbent Assay for Detection of Brucella melitensis-Specific Antibodies in Goat Milk

N. D. Funk,1 L. B. Tabatabai,2 P. H. Elzer,3 S. D. Hagius,3 B. M. Martin,4* and L. J. Hoffman5

Department of Veterinary Microbiology and Preventive Medicine,1 Veterinary Diagnostic Laboratory, College of Veterinary Medicine, Iowa State University,5 Agricultural Research Service, National Animal Disease Center,2 National Veterinary Services Laboratories, U.S. Department of Agriculture, Ames, Iowa,4 Department of Veterinary Science, Louisiana State University AgCenter, Baton Rouge, Louisiana3

Received 19 March 2004/ Returned for modification 10 June 2004/ Accepted 7 August 2004

Brucella melitensis is the cause of brucellosis in sheep and goats, which often results in abortion. Few cases of B. melitensis infection in goats have occurred in the United States over the last 25 years. However, vigilance must be maintained, as it is for the bovine milk industry, to ensure that brucellosis is not introduced into the U.S. goat population. The objective of this study was to develop a sensitive and specific indirect enzyme-linked immunosorbent assay (iELISA) for the detection of B. melitensis-specific antibodies in goat milk. Brucella salt-extractable protein extract was employed as an antigen, and a horseradish peroxidase-labeled polyclonal anti-goat antibody was used as an anti-species conjugate. Thirteen of 13 (100%) individual infected goat milk samples tested positive and 134 of 134 (100%) uninfected bulk milk samples tested negative by the developed iELISA. Three positive milk samples with high, medium, and low absorbance values were used to simulate one positive animal in an otherwise negative herd. By this estimation, one high-titer animal could be detected in a herd of >1,600 animals. Detection estimates for medium- and low-titer animals were one positive animal per herd of <200 and 50 animals, respectively. Based on this estimation, it is recommended that herds be sampled in groups of 50 animals or less for bulk milk testing. The iELISA developed for this study was found to be sensitive and specific and shows potential for use as a bulk milk test for the detection of B. melitensis-specific antibodies in goat milk.

* Corresponding author. Mailing address: National Veterinary Services Laboratories, U.S. Department of Agriculture, 1800 Dayton Ave., Ames, IA 50010. Phone: (515) 663-7731. Fax: (515) 663-7397. E-mail: Barbara.M.Martin{at}aphis.usda.gov.

Journal of Clinical Microbiology, February 2005, p. 721-725, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.721-725.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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