title: Validation of a competitive enzyme-linked immunosorbent assay for detection of antibodies against Babesia bovis.
abstract: 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.
authors.affiliation: Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630, USA. wgoff@vetmed.wsu.edu
authors.forename: Will L
authors.initials: WL
authors.lastname: Goff
authors.forename: John B
authors.initials: JB
authors.lastname: Molloy
authors.forename: Wendell C
authors.initials: WC
authors.lastname: Johnson
authors.forename: Carlos E
authors.initials: CE
authors.lastname: Suarez
authors.forename: Ignacio
authors.initials: I
authors.lastname: Pino
authors.forename: Abdelkebir
authors.initials: A
authors.lastname: Rhalem
authors.forename: Hamid
authors.initials: H
authors.lastname: Sahibi
authors.forename: Luigi
authors.initials: L
authors.lastname: Ceci
authors.forename: Grazia
authors.initials: G
authors.lastname: Carelli
authors.forename: D Scott
authors.initials: DS
authors.lastname: Adams
authors.forename: Travis C
authors.initials: TC
authors.lastname: McGuire
authors.forename: Donald P
authors.initials: DP
authors.lastname: Knowles
authors.forename: Terry F
authors.initials: TF
authors.lastname: McElwain
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000913:Antibodies, Protozoan / Q000097:blood; D016793:Babesia bovis / Q000235:genetics / Q000276:immunology*; D001404:Babesiosis / Q000276:immunology*; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan