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Title: Specific detection of Mycobacterium paratuberculosis DNA associated with granulomatous tissue in Crohn's disease [see comments]
Title Abreviation: Gut Date of Pub: 1994 Apr
Author: Fidler HM; Thurrell W; Johnson NM; Rook GA; McFadden JJ;
Issue/Part/Supplement: 4 Volume Issue: 35 Pagination: 506-10
MESH Headings: Adult; Base Sequence; Blotting, Southern; Crohn Disease (*MI); Double-Blind Method; DNA, Bacterial (*AN); Female; Granuloma (MI); Human; Intestinal Diseases (MI); Intestines (*MI); Male; Middle Age; Molecular Sequence Data; Mycobacterium paratuberculosis (*IP); Polymerase Chain Reaction; Sensitivity and Specificity; Support, Non-U.S. Gov't; -RN-;
Journal Title Code: FVT Publication Type: JOURNAL ARTICLE
Date of Entry: 940607NEntry Month: 9408
Country: ENGLAND Index Priority: 1
Language: Eng Unique Identifier: 94229581
Unique Identifier: 94229581 ISSN: 0017-5749
Abstract: The role of mycobacteria, specifically Mycobacterium paratuberculosis, in Crohn's disease has aroused considerable controversy for many years. Using the ultra sensitive polymerase chain reaction some studies have reported detection of M paratuberculosis DNA in as many as 65% of Crohn's disease patients but also in patients without disease. Other studies have been negative for both groups. We therefore designed a double blind control trial to investigate the presence of mycobacterial DNA in age, sex, and tissue matched paraffin wax embedded tissues from 31 Crohn's disease tissues, 20 diseased gut control tissues, and 10 ulcerative colitis tissues. The specimens were coded and analysed blind with three separate polymerase chain reactions (PCR) based on DNA sequences specific for M paratuberculosis (IS900), M avium (RFLP type A/1) (IS901), and the Mycobacterium genus (65 kDa gene, TB600). The number of granulomata and presence of acid fast bacilli in each Crohn's disease tissue was also investigated. The sensitivity of the system was determined using similarly prepared gut tissue from an animal infected with M paratuberculosis. Four of 31 Crohn's disease tissues and none of the 30 control and ulcerative colitis derived tissues amplified M paratuberculosis DNA. Crohn's disease tissues containing granulomata were significantly more likely to amplify M paratuberculosis specific DNA on PCR than the non-Crohn's disease tissues (p = 0.02). All the positive Crohn's disease tissues contained granulomata, and none contained acid fast bacilli. Equivalent numbers of Crohn's and non-Crohn's disease tissues amplified the region of the 65 kD gene on PCR for non-specific mycobacterial DNA (11/31 and 9/30 respectively). No sections produced an amplified product with the IS901 PCR.(ABSTRACT TRUNCATED AT 250 WORDS)
Abstract By: Author
Address: Department of Medicine, University College London Medical School, Guildford.
Last Revision Date: 951016