Skip to main content
Canadian Journal of Veterinary Research logoLink to Canadian Journal of Veterinary Research
. 1992 Apr;56(2):148–153.

Evaluation of conventional and radiometric fecal culture and a commercial DNA probe for diagnosis of Mycobacterium paratuberculosis infections in cattle.

D C Sockett 1, D J Carr 1, M T Collins 1
PMCID: PMC1263523  PMID: 1591658

Abstract

Radiometric (RCM) and conventional fecal culture (HEY) and a commercial polymerase chain reaction/DNA probe were evaluated as diagnostic tests for subclinical paratuberculosis in dairy cattle using fecal specimens from a repository of paratuberculosis specimens. The case definition of subclinical bovine paratuberculosis was isolation of Mycobacterium paratuberculosis, by conventional or radiometric culture, from fecal samples or internal organs of dairy cattle without diarrhea or chronic weight loss. Animals designated as free of the disease originated exclusively from certified paratuberculosis-free herds in Wisconsin. Among 182 infected cattle, RCM and HEY fecal culture and the DNA probe had test sensitivities of 54.4%, 45.1% and 33.5%, respectively. Fecal samples from only 111 of the M. paratuberculosis-infected cows tested positive by at least one of the three tests and these cows were designated as fecal shedders; the remaining 71 were considered to have prepatent infections. Among the 111 M. paratuberculosis fecal shedders, RCM, HEY and the probe detected the organism in 89.2%, 73.8% and 55.0% of the fecal specimens, respectively. Herd prevalence significantly affected the sensitivity of all three diagnostic tests (p less than 0.05) but only affected the fecal shedder detection efficiency of the DNA probe (p less than 0.01). No positive DNA probe results were found on 100 randomly selected fecal samples from cows in four certified paratuberculosis-free herds, thus the DNA probe was 100% specific. Probe analyses could be performed in 24 h or less. Time to complete the culture-based tests was 12 wk for HEY and 7 wk for RCM.(ABSTRACT TRUNCATED AT 250 WORDS)

Full text

PDF
148

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chiodini R. J., Van Kruiningen H. J., Merkal R. S. Ruminant paratuberculosis (Johne's disease): the current status and future prospects. Cornell Vet. 1984 Jul;74(3):218–262. [PubMed] [Google Scholar]
  2. Colgrove G. S., Thoen C. O., Blackburn B. O., Murphy C. D. Paratuberculosis in cattle: a comparison of three serologic tests with results of fecal culture. Vet Microbiol. 1989 Feb;19(2):183–187. doi: 10.1016/0378-1135(89)90083-7. [DOI] [PubMed] [Google Scholar]
  3. Collins M. T., Kenefick K. B., Sockett D. C., Lambrecht R. S., McDonald J., Jorgensen J. B. Enhanced radiometric detection of Mycobacterium paratuberculosis by using filter-concentrated bovine fecal specimens. J Clin Microbiol. 1990 Nov;28(11):2514–2519. doi: 10.1128/jcm.28.11.2514-2519.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hurley S. S., Splitter G. A., Welch R. A. Development of a diagnostic test for Johne's disease using a DNA hybridization probe. J Clin Microbiol. 1989 Jul;27(7):1582–1587. doi: 10.1128/jcm.27.7.1582-1587.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kim Y. G., Bech-Nielsen S., Gordon J. C., Slemons R. D., Spangler E. Comparison of two methods for isolation of Mycobacterium paratuberculosis from bovine fecal samples. Am J Vet Res. 1989 Jul;50(7):1110–1113. [PubMed] [Google Scholar]
  6. Larsen A. B., Merkal R. S., Cutlip R. C. Age of cattle as related to resistance to infection with Mycobacterium paratuberculosis. Am J Vet Res. 1975 Mar;36(3):255–257. [PubMed] [Google Scholar]
  7. Ransohoff D. F., Feinstein A. R. Problems of spectrum and bias in evaluating the efficacy of diagnostic tests. N Engl J Med. 1978 Oct 26;299(17):926–930. doi: 10.1056/NEJM197810262991705. [DOI] [PubMed] [Google Scholar]
  8. Rogan W. J., Gladen B. Estimating prevalence from the results of a screening test. Am J Epidemiol. 1978 Jan;107(1):71–76. doi: 10.1093/oxfordjournals.aje.a112510. [DOI] [PubMed] [Google Scholar]
  9. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  10. Seitz S. E., Heider L. E., Heuston W. D., Bech-Nielsen S., Rings D. M., Spangler L. Bovine fetal infection with Mycobacterium paratuberculosis. J Am Vet Med Assoc. 1989 May 15;194(10):1423–1426. [PubMed] [Google Scholar]
  11. Sherman D. M., Bray B., Gay J. M., Bates F. Evaluation of the agar gel immunodiffusion test for diagnosis of subclinical paratuberculosis in cattle. Am J Vet Res. 1989 Apr;50(4):525–530. [PubMed] [Google Scholar]
  12. Sherman D. M., Gay J. M., Bouley D. S., Nelson G. H. Comparison of the complement-fixation and agar gel immunodiffusion tests for diagnosis of subclinical bovine paratuberculosis. Am J Vet Res. 1990 Mar;51(3):461–465. [PubMed] [Google Scholar]
  13. Sherman D. M., Markham R. J., Bates F. Agar gel immunodiffusion test for diagnosis of clinical paratuberculosis in cattle. J Am Vet Med Assoc. 1984 Jul 15;185(2):179–182. [PubMed] [Google Scholar]
  14. Vary P. H., Andersen P. R., Green E., Hermon-Taylor J., McFadden J. J. Use of highly specific DNA probes and the polymerase chain reaction to detect Mycobacterium paratuberculosis in Johne's disease. J Clin Microbiol. 1990 May;28(5):933–937. doi: 10.1128/jcm.28.5.933-937.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Vecchio T. J. Predictive value of a single diagnostic test in unselected populations. N Engl J Med. 1966 May 26;274(21):1171–1173. doi: 10.1056/NEJM196605262742104. [DOI] [PubMed] [Google Scholar]
  16. Yokomizo Y., Yugi H., Merkal R. S. A method for avoiding false-positive reactions in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of bovine paratuberculosis. Nihon Juigaku Zasshi. 1985 Feb;47(1):111–119. doi: 10.1292/jvms1939.47.111. [DOI] [PubMed] [Google Scholar]

Articles from Canadian Journal of Veterinary Research are provided here courtesy of Canadian Veterinary Medical Association

RESOURCES