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Comparative Study
. 2005 Jun;12(6):685-92.
doi: 10.1128/CDLI.12.6.685-692.2005.

Evaluation of five antibody detection tests for diagnosis of bovine paratuberculosis

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Comparative Study

Evaluation of five antibody detection tests for diagnosis of bovine paratuberculosis

Michael T Collins et al. Clin Diagn Lab Immunol. 2005 Jun.

Abstract

Five diagnostic tests based on enzyme-linked immunosorbent assay (ELISA) technology for bovine paratuberculosis were evaluated by using individual serum or milk samples from 359 dairy cattle in seven paratuberculosis-free herds and 2,094 dairy cattle in seven Mycobacterium paratuberculosis-infected dairy herds. Three independent laboratories using three different culture procedures completed fecal cultures for M. paratuberculosis on these cattle and found 417 cows to be shedding M. paratuberculosis in their feces. An animal that was fecal culture positive for M. paratuberculosis by any of the three laboratories was considered a confirmed case of infection. The specificity of three ELISAs (two on serum and one on milk) was > or =99.8%. The specificity of the remaining two ELISAs, both done on serum, was 94.9 and 84.7%. Four of the five ELISAs evaluated produced similar sensitivity in detecting fecal culture-positive cattle (27.8 to 28.9%). Serum ELISA "D" had the lowest specificity (84.7%) and the highest sensitivity (44.5%), but if the cutoff value defining a positive test was changed from 125 to 250% (of the positive control) the sensitivity and specificity, 31.8 and 97.5%, respectively, were comparable to those of the other four assays. If the case definition for M. paratuberculosis infection was based on the culture results of a single laboratory instead of the combined results of three laboratories, ELISA sensitivity estimates were 45.7 to 50.0%. With the exception of ELISA D, assay agreement was high (kappa 0.66 to 0.85) for categorical assay interpretations (positive or negative), but linear regression of quantitative results showed low correlation coefficients (r(2) = 0.40 to 0.68) due to the fact that ELISA results for some cows were high in one assay but low in another assay. Likelihood ratio analysis showed a direct relationship between the magnitude of ELISA result and the odds of a cow shedding M. paratuberculosis in its feces. If used judiciously and interpreted quantitatively, these ELISAs are useful tools in support of paratuberculosis control programs in dairy herds.

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Figures

FIG. 1.
FIG. 1.
Number and percentage of cows at each level of fecal shedding based on their average fecal shedding score for 443 M. paratuberculosis-infected cows among seven infected herds.
FIG. 2.
FIG. 2.
Scatter plots with linear regression lines comparing quantitative results for four pairs of ELISAs selected to show a range of correlations. (A) Kit A and kit B; (B) kit A and kit C; (C) kit D and kit E; (D) kit B and kit E.

References

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    1. Collins, D. M., D. M. Gabric, and G. W. de Lisle. 1989. Identification of a repetitive DNA sequence specific to Mycobacterium paratuberculosis. FEMS Microbiol. Lett. 60:175-178. - PubMed
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    1. Collins, M. T. 2002. Interpretation of a commercial bovine paratuberculosis enzyme-linked immunosorbent assay by using likelihood ratios. Clin. Diagn. Lab. Immunol. 9:1367-1371. - PMC - PubMed
    1. Collins, M. T., K. B. Kenefick, D. C. Sockett, R. S. Lambrecht, J. McDonald, and J. B. Jørgensen. 1990. Enhanced radiometric detection of Mycobacterium paratuberculosis using filter concentrated fecal specimens. J. Clin. Microbiol. 28:2514-2519. - PMC - PubMed

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