Polymerase chain reaction detection of Mycobacterium paratuberculosis and Mycobacterium avium subsp silvaticum in long term cultures from Crohn's disease and control tissues.

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Title: Polymerase chain reaction detection of Mycobacterium paratuberculosis and Mycobacterium avium subsp silvaticum in long term cultures from Crohn's disease and control tissues.
Title Abreviation: Gut		Date of Pub: 1992 Sep
Author: Moss MT; Sanderson JD; Tizard ML; Hermon-Taylor J;
Issue/Part/Supplement: 9	Volume Issue: 33		Pagination: 1209-13
MESH Headings: Base Sequence; Colitis, Ulcerative (MI); Crohn Disease (MI); Human; Molecular Sequence Data; Mycobacterium avium (IP); Mycobacterium paratuberculosis (*IP); Polymerase Chain Reaction; Support, Non-U.S. Gov't; Time Factors; Tissue Culture; -AA-;
Journal Title Code: FVT	Publication Type: JOURNAL ARTICLE
Date of Entry: 921203N	Entry Month: 9302
Country: ENGLAND	Index Priority: 1
Language: Eng	Unique Identifier: 93051661
Unique Identifier: 93051661	ISSN: 0017-5749
Abstract: Thirty one cultures were established in MG3 medium from the intestinal tissues of 29 patients, including 18 with Crohn's disease, five with ulcerative colitis, and six non-inflammatory bowel disease controls. All cultures grew either acid fast bacilli or uncharacterized spheroplasts. Pellets from these cultures were coded and assayed blind for M paratuberculosis and M avium subsp silvaticum using IS900- and IS902-PCR (polymerase chain reaction) assays, respectively. IS900 and IS902 are multicopy DNA insertion elements specific for these two organisms. Six Crohn's disease cultures and a single non-inflammatory bowel disease control were positive for M paratuberculosis. A further six cultures were positive for M avium subsp silvaticum, of which two each were from Crohn's disease, ulcerative colitis, and non-inflammatory bowel disease controls. The intensity of the IS900-PCR signals indicated very low numbers of M paratuberculosis organisms and bore no relation to visible spheroplastic or bacillary mycobacterial growth. The results suggest that M paratuberculosis isolated from man exists in a form which hardly replicates if at all when cultured in MG3 medium in vitro, and are consistent with the involvement of this known animal enteric pathogen in a proportion of chronic enteritis in man.
Abstract By: Author
Address: Department of Surgery, St George's Hospital Medical School, London.