Validation of a Competitive ELISA for Detection of Babesia bigemina Antibodies in Cattle

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Validation of a Competitive ELISA for Detection of Babesia bigemina Antibodies in Cattle.

Clin Vaccine Immunol. 2008 Jul 16;

Authors: Goff WL, Johnson WC, Molloy JB, Jorgensen WK, Waldron SJ, Figueroa JV, Matthee O, Adams DS, McGuire TC, Pino I, Mosqueda J, Palmer GH, Suarez CE, Knowles DP, McElwain TF

A competitive ELISA (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 (ROC) analysis revealed 16% inhibition as the threshold for a negative sample with an associated specificity of 98.3% and sensitivity of 94.7%. Increasing the threshold to 21% increased the specificity to 100% with a modest decrease in sensitivity to 87.2%. Using 21% inhibition, positive predictive values ranged from 90.7% (10% prevalence) to 100% (95% prevalence) and 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 seven days post-intravenous inoculation. The cELISA was distributed to 5 different laboratories along with a reference set of 100 defined bovine sera including known positive, known negative and field samples. Pairwise concordance among the 5 laboratories ranged from 100% to 97% with Kappa values all above 0.8 indicating a high degree of reliability. Overall, the cELISA appears to have the attributes necessary for international application.

PMID: 18632921 [PubMed - as supplied by publisher]

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