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Validation of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Babesia bigemina Antibodies in Cattle

Goff, W.L. and Johnson, W.C. and Molloy, J.B. and Jorgensen, W.K. and Waldron, S.J. and Figueroa, J.V. and Matthee, O. and Adams, D.S. and McGuire, T.C. and Pino, I. and Mosqueda, J. and Palmer, G.H. and Suarez, C.E. and Knowles, D.P. and McElwain, T.F. (2008) Validation of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Babesia bigemina Antibodies in Cattle. Clinical and Vaccine Immunology, 15 (9). pp. 1316-1321.

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Article Link(s): http://dx.doi.org/10.1128/CVI.00150-08

Publisher URL: http://journals.asm.org

Abstract

A competitive enzyme-linked immunosorbent assay (cELISA) based on a broadly conserved, species-specific, B-cell epitope within the C terminus of Babesia bigemina rhoptry-associated protein 1a was validated for international use. Receiver operating characteristic analysis revealed 16% inhibition as the threshold for a negative result, with an associated specificity of 98.3% and sensitivity of 94.7%. Increasing the threshold to 21% increased the specificity to 100% but modestly decreased the sensitivity to 87.2%. By using 21% inhibition, the positive predictive values ranged from 90.7% (10% prevalence) to 100% (95% prevalence) and the negative predictive values ranged from 97.0% (10% prevalence) to 48.2% (95% prevalence). The assay was able to detect serum antibody as early as 7 days after intravenous inoculation. The cELISA was distributed to five different laboratories along with a reference set of 100 defined bovine serum samples, including known positive, known negative, and field samples. The pairwise concordance among the five laboratories ranged from 100% to 97%, and all kappa values were above 0.8, indicating a high degree of reliability. Overall, the cELISA appears to have the attributes necessary for international application.

Item Type:Article
Additional Information:© American Society for Microbiology.
Keywords:Rhoptry-associated protein 1a; expression; epitopes; Bovis; infection; surface; genes.
Subjects:Science > Science (General)
Veterinary medicine > Veterinary parasitology
Veterinary medicine > Diseases of special classes of animals > Cattle
Deposited On:02 Feb 2009 02:02
Last Modified:20 Apr 2011 23:18

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