title: Mycobacterium paratuberculosis and Mycobacterium avium subsp. silvaticum DNA cannot be detected by PCR in Crohn's disease tissue.
abstract: BACKGROUND
  The etiology of Crohn's disease remains unknown. A putative mycobacterial cause of the disease is still controversial.
  
  
  OBJECTIVE
  To assess the mycobacterial hypothesis in Crohn's disease using a polymerase chain reaction technique.
  
  
  METHODS
  Nested polymerase chain reaction with primers on the 16S-rRNA coding region (16S-rDNA) and with primers specific both to the insertion sequences (IS) 900, and IS 901/902 were used to amplify Mycobacterium paratuberculosis or Mycobacterium avium subsp. silvaticum DNA in frozen endoscopic intestinal biopsies or surgical resection specimens from patients with Crohn's disease (n = 47: 25 endoscopic biopsies and 22 surgical resection samples, +/- lymph nodes), ulcerative colitis (n = 27), and non inflammatory bowel diseases (n = 20: colonic tumors and diverticulitis). Positive as well as negative controls were used throughout the study.
  
  
  RESULTS
  All strains of Mycobacterium paratuberculosis and Mycobacterium avium subsp. silvaticum tested were positive for both primer systems. Of the 94 biopsies tested, 5 (2 Crohn's disease, 1 ulcerative colitis and 2 controls) were positive with the 16S-rDNA primers but did not correspond to Mycobacterium paratuberculosis or Mycobacterium avium subsp. silvaticum. None of the specimens was positive with the IS primers.
  
  
  CONCLUSIONS
  These results do not support the hypothesis that Mycobacterium paratuberculosis, or Mycobacterium avium subsp. silvaticum play a role in Crohn's disease.
authors.affiliation: Département de Pathologie Digestive, Hôpital Laennec, Paris.
authors.forename: C
authors.initials: C
authors.lastname: Cellier
authors.forename: H
authors.initials: H
authors.lastname: De Beenhouwer
authors.forename: A
authors.initials: A
authors.lastname: Berger
authors.forename: C
authors.initials: C
authors.lastname: Penna
authors.forename: F
authors.initials: F
authors.lastname: Carbonnel
authors.forename: R
authors.initials: R
authors.lastname: Parc
authors.forename: P H
authors.initials: PH
authors.lastname: Cugnenc
authors.forename: Y
authors.initials: Y
authors.lastname: Le Quintrec
authors.forename: J P
authors.initials: JP
authors.lastname: Gendre
authors.forename: J P
authors.initials: JP
authors.lastname: Barbier
authors.forename: F
authors.initials: F
authors.lastname: Portaels
mesh_terms: D000328:Adult; D003424:Crohn Disease / Q000382:microbiology*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008297:Male; D009162:Mycobacterium avium / Q000235:genetics* / Q000302:isolation & purification; D016927:Mycobacterium avium subsp. paratuberculosis / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction
chemical_list: D004269:DNA, Bacterial