Skip to main content
PMC home page
Gut logoLink to Gut
. 1995 Nov;37(5):660–667. doi: 10.1136/gut.37.5.660

Mycobacterium paratuberculosis DNA not detected in Crohn's disease tissue by fluorescent polymerase chain reaction.

D S Rowbotham 1, N P Mapstone 1, L K Trejdosiewicz 1, P D Howdle 1, P Quirke 1
PMCID: PMC1382871  PMID: 8549942

Abstract

The role of mycobacteria in the aetiology of Crohn's disease has been a contentious subject for many years. Mycobacterium paratuberculosis is known to cause a chronic granulomatous enteritis in animals (Johne's disease) and has been implicated as a possible infectious cause of Crohn's disease. However this fastidious organism is only rarely detected by conventional microbiological techniques. This study used oligonucleotide primers to the species-specific M paratuberculosis IS900 DNA insertion element and the polymerase chain reaction to amplify any M paratuberculosis DNA from intestinal tissue DNA extracts. One oligonucleotide primer was fluorochrome-labelled and the presence of fluorescent amplified product was determined using an automated DNA sequencer with a computerised gel-scanning laser. This method was shown capable of detecting 1-2 mycobacterial genomes. Intestinal tissue samples were obtained from 68 patients with histologically confirmed Crohn's disease, 49 patients with histologically confirmed ulcerative colitis, and 26 non-inflammatory bowel disease controls. In no case was M paratuberculosis detected in any of the inflammatory bowel disease tissue samples and only one non-inflammatory bowel disease case was positive. These results do not support the hypothesis that M paratuberculosis has an aetiological role in Crohn's disease.

Full text

PDF
664

Images in this article

662Figure 1
on p.662
663Figure 3
on p.663
664Figure 4
on p.664

Selected References

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

  1. Chiodini R. J. Crohn's disease and the mycobacterioses: a review and comparison of two disease entities. Clin Microbiol Rev. 1989 Jan;2(1):90–117. doi: 10.1128/cmr.2.1.90. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Chiodini R. J., Van Kruiningen H. J., Merkal R. S., Thayer W. R., Jr, Coutu J. A. Characteristics of an unclassified Mycobacterium species isolated from patients with Crohn's disease. J Clin Microbiol. 1984 Nov;20(5):966–971. doi: 10.1128/jcm.20.5.966-971.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chiodini R. J., Van Kruiningen H. J., Thayer W. R., Merkal R. S., Coutu J. A. Possible role of mycobacteria in inflammatory bowel disease. I. An unclassified Mycobacterium species isolated from patients with Crohn's disease. Dig Dis Sci. 1984 Dec;29(12):1073–1079. doi: 10.1007/BF01317078. [DOI] [PubMed] [Google Scholar]
  5. Dell'Isola B., Poyart C., Goulet O., Mougenot J. F., Sadoun-Journo E., Brousse N., Schmitz J., Ricour C., Berche P. Detection of Mycobacterium paratuberculosis by polymerase chain reaction in children with Crohn's disease. J Infect Dis. 1994 Feb;169(2):449–451. doi: 10.1093/infdis/169.2.449. [DOI] [PubMed] [Google Scholar]
  6. Erlich H. A., Gelfand D., Sninsky J. J. Recent advances in the polymerase chain reaction. Science. 1991 Jun 21;252(5013):1643–1651. doi: 10.1126/science.2047872. [DOI] [PubMed] [Google Scholar]
  7. Fidler H. M., Thurrell W., Johnson N. M., Rook G. A., McFadden J. J. Specific detection of Mycobacterium paratuberculosis DNA associated with granulomatous tissue in Crohn's disease. Gut. 1994 Apr;35(4):506–510. doi: 10.1136/gut.35.4.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gitnick G., Collins J., Beaman B., Brooks D., Arthur M., Imaeda T., Palieschesky M. Preliminary report on isolation of mycobacteria from patients with Crohn's disease. Dig Dis Sci. 1989 Jun;34(6):925–932. doi: 10.1007/BF01540280. [DOI] [PubMed] [Google Scholar]
  9. Green E. P., Tizard M. L., Moss M. T., Thompson J., Winterbourne D. J., McFadden J. J., Hermon-Taylor J. Sequence and characteristics of IS900, an insertion element identified in a human Crohn's disease isolate of Mycobacterium paratuberculosis. Nucleic Acids Res. 1989 Nov 25;17(22):9063–9073. doi: 10.1093/nar/17.22.9063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hermon-Taylor J., Moss M., Tizard M., Malik Z., Sanderson J. Molecular biology of Crohn's disease mycobacteria. Baillieres Clin Gastroenterol. 1990 Mar;4(1):23–42. doi: 10.1016/0950-3528(90)90037-h. [DOI] [PubMed] [Google Scholar]
  11. McClure H. M., Chiodini R. J., Anderson D. C., Swenson R. B., Thayer W. R., Coutu J. A. Mycobacterium paratuberculosis infection in a colony of stumptail macaques (Macaca arctoides). J Infect Dis. 1987 May;155(5):1011–1019. doi: 10.1093/infdis/155.5.1011. [DOI] [PubMed] [Google Scholar]
  12. McCord C., Freeman H. P. Excess mortality in Harlem. N Engl J Med. 1990 Jan 18;322(3):173–177. doi: 10.1056/NEJM199001183220306. [DOI] [PubMed] [Google Scholar]
  13. McFadden J. J., Butcher P. D., Chiodini R., Hermon-Taylor J. Crohn's disease-isolated mycobacteria are identical to Mycobacterium paratuberculosis, as determined by DNA probes that distinguish between mycobacterial species. J Clin Microbiol. 1987 May;25(5):796–801. doi: 10.1128/jcm.25.5.796-801.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moss M. T., Green E. P., Tizard M. L., Malik Z. P., Hermon-Taylor J. Specific detection of Mycobacterium paratuberculosis by DNA hybridisation with a fragment of the insertion element IS900. Gut. 1991 Apr;32(4):395–398. doi: 10.1136/gut.32.4.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Portaels F., Larsson L., Smeets P. Isolation of mycobacteria from healthy persons' stools. Int J Lepr Other Mycobact Dis. 1988 Sep;56(3):468–471. [PubMed] [Google Scholar]
  17. Sanderson J. D., Moss M. T., Tizard M. L., Hermon-Taylor J. Mycobacterium paratuberculosis DNA in Crohn's disease tissue. Gut. 1992 Jul;33(7):890–896. doi: 10.1136/gut.33.7.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thorel M. F., Krichevsky M., Lévy-Frébault V. V. Numerical taxonomy of mycobactin-dependent mycobacteria, emended description of Mycobacterium avium, and description of Mycobacterium avium subsp. avium subsp. nov., Mycobacterium avium subsp. paratuberculosis subsp. nov., and Mycobacterium avium subsp. silvaticum subsp. nov. Int J Syst Bacteriol. 1990 Jul;40(3):254–260. doi: 10.1099/00207713-40-3-254. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Wu S. W., Pao C. C., Chan J., Yen T. S. Lack of mycobacterial DNA in Crohn's disease tissue. Lancet. 1991 Jan 19;337(8734):174–175. doi: 10.1016/0140-6736(91)90837-f. [DOI] [PubMed] [Google Scholar]

Articles from Gut are provided here courtesy of BMJ Publishing Group

RESOURCES