title: Polymerase chain reaction detection of Mycobacterium paratuberculosis and Mycobacterium avium subsp silvaticum in long term cultures from Crohn's disease and control tissues.
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.
authors.affiliation: Department of Surgery, St George's Hospital Medical School, London.
authors.forename: M T
authors.initials: MT
authors.lastname: Moss
authors.forename: J D
authors.initials: JD
authors.lastname: Sanderson
authors.forename: M L
authors.initials: ML
authors.lastname: Tizard
authors.forename: J
authors.initials: J
authors.lastname: Hermon-Taylor
authors.forename: F A
authors.initials: FA
authors.lastname: el-Zaatari
authors.forename: D C
authors.initials: DC
authors.lastname: Markesich
authors.forename: D Y
authors.initials: DY
authors.lastname: Graham
mesh_terms: D001483:Base Sequence; D003093:Colitis, Ulcerative / Q000382:microbiology; D003424:Crohn Disease / Q000382:microbiology*; D046508:Culture Techniques; D006801:Humans; D008969:Molecular Sequence Data; D009162:Mycobacterium avium / Q000302:isolation & purification*; D016927:Mycobacterium avium subsp. paratuberculosis / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D013997:Time Factors