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Clinical and Vaccine Immunology, November 2006, p. 1212-1216, Vol. 13, No. 11
1071-412X/06/$08.00+0     doi:10.1128/CVI.00196-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Validation of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Babesia bovis{triangledown}

Will L. Goff,1* John B. Molloy,2 Wendell C. Johnson,1 Carlos E. Suarez,1 Ignacio Pino,3 Abdelkebir Rhalem,4 Hamid Sahibi,4 Luigi Ceci,5 Grazia Carelli,5 D. Scott Adams,6 Travis C. McGuire,7 Donald P. Knowles,1 and Terry F. McElwain7,8

Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630,1 Queensland Department of Primary Industries, Animal Research Institute, Yeerongpilly, Queensland, 4105, Australia,2 Veterinary Clinic, Route 349 KM1.0 Interior, Mayaguez, Puerto Rico 00680,3 Department de Parasitologie, Institut Agronomique et Veterinaire Hassan II, 6202 Rabat, Morocco,4 Division of Clinical Veterinary Medicine, Department of Animal Health and Welfare, Faculty of Veterinary Medicine—University of Bari, Strada per Casamassima km 3-70010 Valenzano (Ba.), Italy,5 VMRD, Inc., Pullman, Washington 99163,6 Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, Washington 99164-7040,7 Washington Animal Disease Diagnostic Laboratory, Pullman, Washington 99163-20378

Received 30 May 2006/ Returned for modification 11 August 2006/ Accepted 28 August 2006

A previously developed competitive enzyme-linked immunosorbent assay (cELISA) based on a species-specific, broadly conserved, and tandemly repeated B-cell epitope within the C terminus of rhoptry-associated protein 1 of Babesia bovis was refined and validated for use internationally. Receiver operating characteristic analysis revealed an assay with a specificity and positive predictive value of 100% and a sensitivity of 91.1%, with various negative predictive values depending on the level of disease prevalence. The cELISA was distributed to four different laboratories, along with a reference set of 100 defined bovine sera, including known-positive, known-negative, and field samples. Pairwise concordances among the four laboratories ranged from 94% to 88%. Analysis of variance of the resulting optical densities and a test of homogeneity indicated no significant difference among the laboratories. Overall, the cELISA appears to have the attributes necessary for international application.

* Corresponding author. Mailing address: Animal Disease Research Unit, USDA-ARS, Washington State University, Pullman, WA 99164-6630. Phone: (509) 335-6003. Fax: (509) 335-8328. E-mail: wgoff{at}vetmed.wsu.edu.

{triangledown} Published ahead of print on 6 September 2006.

Clinical and Vaccine Immunology, November 2006, p. 1212-1216, Vol. 13, No. 11
1071-412X/06/$08.00+0     doi:10.1128/CVI.00196-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Goff, W. L., Johnson, W. C., Molloy, J. B., Jorgensen, W. K., Waldron, S. J., Figueroa, J. V., Matthee, O., Adams, D. S., McGuire, T. C., Pino, I., Mosqueda, J., Palmer, G. H., Suarez, C. E., Knowles, D. P., McElwain, T. F. (2008). Validation of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Babesia bigemina Antibodies in Cattle. CVI 15: 1316-1321 [Abstract] [Full Text]  
  • Howell, J. M., Ueti, M. W., Palmer, G. H., Scoles, G. A., Knowles, D. P. (2007). Persistently Infected Calves as Reservoirs for Acquisition and Transovarial Transmission of Babesia bovis by Rhipicephalus (Boophilus) microplus. J. Clin. Microbiol. 45: 3155-3159 [Abstract] [Full Text]  


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