Validation of a competitive enzyme-linked immunosorbent assay for detection of antibodies against Babesia bovis.
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.
Goff
Will L
WL
Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630, USA. wgoff@vetmed.wsu.edu
Molloy
John B
JB
Johnson
Wendell C
WC
Suarez
Carlos E
CE
Pino
Ignacio
I
Rhalem
Abdelkebir
A
Sahibi
Hamid
H
Ceci
Luigi
L
Carelli
Grazia
G
Adams
D Scott
DS
McGuire
Travis C
TC
Knowles
Donald P
DP
McElwain
Terry F
TF
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
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan