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Validation of a competitive enzyme-linked immunosorbent assay for detection of Babesia bigemina antibodies in cattle.
Title | Validation of a competitive enzyme-linked immunosorbent assay for detection of Babesia bigemina antibodies in cattle. |
Publication Type | Journal Article |
Year of Publication | 2008 |
Authors | Goff, WL, Johnson, WC, Molloy, JB, Jorgensen, WK, Waldron, SJ, Figueroa, JV, Matthee, O, D Adams, S, McGuire, TC, Pino, I, Mosqueda, J, Palmer, GH, Suarez, CE, Knowles, DP, McElwain, TF |
Journal | Clin Vaccine Immunol |
Volume | 15 |
Issue | 9 |
Pagination | 1316-21 |
Date Published | 2008 Sep |
ISSN | 1556-679X |
Keywords | Animals, Antibodies, Protozoan, Babesia, Babesiosis, Cattle, Cattle Diseases, Enzyme-Linked Immunosorbent Assay, Predictive Value of Tests, Protozoan Proteins, ROC Curve, Sensitivity and Specificity, Time Factors |
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. |
DOI | 10.1128/CVI.00150-08 |
Alternate Journal | Clin. Vaccine Immunol. |
PubMed ID | 18632921 |
PubMed Central ID | PMC2546671 |