Diagnostic performance of enzyme-linked immnosorbent assays for diagnosing paratuberculosis in cattle: a meta-analysis

  • Pak, Son-Il (Department of Veterinary Medicine, Kangwon National University)
  • Accepted : 2004.08.23
  • Published : 2004.12.30


To evaluate the diagnostic accuracy of two commercial ELISA tests (Allied- and CSL-ELISA) for the diagnosis of Mycobacterium paratuberculosis in cattle, Meta-analysis using English language papers published during 1990-2001 was performed. Diagnostic odds ratios (DOR) were analyzed using regression analysis together with summary receiver operating characteristic (ROC) curves. The difference in diagnostic performance between the two ELISA systems was evaluated by using linear regression. Publication bias was assessed by funnel plot and linear regression. The pooled sensitivity and specificity were 44% (95% CI, 38 to 51) and 98% (95% CI, 96 to 99) for the random-effect model. The DOR between studies was heterogeneous. The area under the fitted ROC curve (AUC) was 0.72 for the unweighted and 0.77 for the weighted model. Maximum joint sensitivity and specificity for the unweighted and weighted model from their summary ROC curve were 70% and 75%, respectively. Based on the fitted model, at a specificity of 95%, sensitivity was estimated to be 52% for the unweighted and 57% for the weighted model. From the final multivariable model study characteristic, the country was the only significant variable with an explained component variance of 13.3%. There were no significant differences in discriminatory power, sensitivity, and specificity between the two ELISA tests. The overall diagnostic accuracy of two commercial ELISA tests was moderate, as judged by the AUC, maximum joint sensitivity and specificity, and estimates from the fitted model and clinical usefulness of the tests for screening program is limited because of low sensitivity and heterogeneous of DOR. It is, therefore, recommended to use ELISA tests as a parallel testing with other diagnostic tests together to increase test sensitivity in the screening program.


  1. Collins, M. T., Sockett, D. C., Goodger, W. J., Conrad, T. A., Thomas, C. B. and Carr, D. J. Herd prevalence and geographic distribution of, and risk factors for, bovine paratuberculosis in Wisconsin. J. Am. Vet. Med. Assoc. 1994, 204, 636-641
  2. Diamond, G. A. Limited assurance. Am. J. Cardiol. 1989, 63, 99-100
  3. Egger, M., Davey-Smith, G., Schneider, M. and Minder, C. Bias in meta-analysis detected by a single, graphical test. Brit. Med. J. 1997, 315, 629-634
  4. Hedges L. V. Fixed effect models. In: Cooper H, Hedges LV. The handbook of research synthesis, pp. 285-299. Russell Sage Foundation, New York, 1994
  5. Kahn, H. A. and Sempos, C. T. Statistical methods in epidemiology. Oxford University Press, New York, 1989
  6. Littenberg, B. and Moses, L. E. Estimating diagnostic accuracy from multiple conflicting reports: a new metaanalytic method. Med. Decis. Making. 1993, 13, 313-321
  7. Owens, D. K., Holodniy, M., McDonald, T. W., Scott, J. and Sonnad, S. A meta-analytic evaluation of the polymerase chain reaction for the diagnosis of HIVinfection in infants. JAMA. 1996, 275, 1342-1348
  8. Yokomizo, Y., Kishima, M., Mori, Y. and Nishimori, K. Evaluation of enzyme-linked immunosorbent assay in comparison with complement fixation test for thediagnosis of subclinical paratuberculosis in cattle. J. Vet. Med. Sci. 1991, 53, 577-584
  9. Collins, M. T., Sockett, D. C., Ridge, S. and Cox, J. C. Evaluation of a commercial enzyme-linked immunosorbent assay for Johne’s disease. J. Clin.Microbiol. 1991, 29, 272-276
  10. Ridge, S. E., Morgan, I. R., Sockett, D. C., Collins, M.T., Condron, R. J., Skilbeck, N. W. and Webber, J. J. Comparison of the Johne’s absorbed EIA andcomplement fixation test for diagnosis of Johne's disease in cattle. Aust. Vet. J. 1991, 68, 253-257
  11. Sockett, D. C., Conrad, T. A., Thomas, C. B. and Collins, M. T. Evaluation of four serological tests for bovine paratuberculosis. J. Clin. Microbiol. 1992, 30, 1134-1139
  12. Sockett, D. C., Carr, D. J., Richards, W. D. and Collins, M. T. A repository of specimens for comparison of diagnostic testing procedures for bovineparatuberculosis. J. Vet. Diagn. Invest. 1992, 4, 188-191
  13. Cochran, W. G. The combination of estimates from different experiments. Biometrics. 1954, 10, 101-129
  14. Midgette, A. S., Stukel, T. A. and Littenberg. B. A meta-analytic method for summarizing diagnostic test performances: receiver-operating-characteristic-summary point estimates. Med. Decis. Making. 1993, 13, 253-257
  15. Sweeney, R. W., Whitlock, R. H., Buckley, C. L. and Spencer, P. A. Evaluation of a commercial enzymelinked immunosorbent assay for the diagnosis of paratuberculosis in dairy cattle. J. Vet. Diagn. Invest. 1995, 7, 488-493
  16. Swets, J. A. Form of empirical ROCs in discrimination and diagnostic tasks: Implications for theory and measurement of performance. Psychol. Bullet. 1986,99, 181-198
  17. Muskens, J., Barkema, H. W., Russchen, E., van Maanen, K., Schukken, Y. H. and Bakker, D. Prevalence and regional distribution of paratuberculosis in dairy herds in The Netherlands. Vet. Microbiol. 2000, 77, 253-261
  18. Ferrer, R. L. Graphical methods for detecting bias in meta-analysis. Fam. Med. 1998, 30, 579-583
  19. Milner, A. R., Mack, W. N., Coates, K. J., Hill, J., Gill, I. and Sheldrick, P. The sensitivity and specificity of a modified ELISA for the diagnosis of Johne's disease from a field trial in cattle. Vet. Microbiol. 1990, 25, 193-198
  20. DerSimonian, R. and Laird, N. Meta analysis in clinical trials. Control Clin. Trials. 1986, 7, 177-188
  21. Hanley, J. A. and McNeil, B. J. The meaning and use of the area under an ROC curve. Radiology. 1982, 143, 29-36
  22. Moses, L. E., Shapiro, D. and Littenberg. B. Combining independent studies of a diagnostic test into a summary ROC curve: data-analytic approaches andsome additional considerations. Stat. Med. 1993, 12, 1293-1316
  23. Tang, J. L. and Liu, J. L. Y. Misleading funnel plot for detection of bias in meta-analysis. J. Clin. Epidemiol. 2000, 53, 477-484
  24. Turner, R. M., Omar, R. Z., Yang, M., Goldstein, H. and Thompson, S. G. A multilevel model framework for meta-analysis of clinical trials with binary outcomes. Stat. Med. 2000, 19, 3417-3432
  25. Cox, J. C., Drane, D. P., Jones, S. L., Ridge, S. and Milner, A. R. Development and evaluation of a rapid absorbed enzyme immunoassay test for the diagnosisof Johne’s disease in cattle. Aust. Vet. J. 1991, 68, 157-160
  26. Whitlock, R. H., Wells, S. J., Sweeney, R. W. and van Tiem, J. ELISA and fecal culture for paratuberculosis (Johne’s disease): sensitivity and specificity of each method. Vet. Microbiol. 2000, 77, 387-398
  27. Dargatz, D. A., Byrum, B. A., Barber, L. K., Sweeney, R. W., Whitlock, R. H., Shulaw, W. P., Jacobson, R. H. and Stabel, J.R. Evaluation of a commercial ELISA for diagnosis of paratuberculosis in cattle. J. Am. Vet. Med. Assoc. 2001, 218, 1163-1166
  28. Walter, S. D., Irwig, L. and Glasziou, P. P. Metaanalysis of diagnostic tests with imperfect reference standards. J. Clin. Epidemiol. 1999, 52, 943-951