title: Fecal detection of Mycobacterium avium paratuberculosis using the IS900 DNA sequence in Crohn's disease and ulcerative colitis patients and healthy subjects.
abstract: OBJECTIVE
  Despite the increasing evidence of MAP/DNA isolation in Crohn's disease (CD), its potential pathogenetic role remains unclear. To further clarify the possible relationship between MAP and CD, we investigated the presence of IS900 DNA fragment in feces from Crohn's disease and ulcerative colitis (UC) patients and from healthy controls (HC).
  
  
  METHODS
  Stool samples were collected from 31 CD, 20 UC, and 23 HC and stored at -20°C in 200-mg aliquots. DNA was extracted. MAP presence was detected with a specific PCR amplifying a 409-bp fragment from IS900. The specificity of PCR for IS900 was confirmed sequencing three positive products. Statistical analysis was performed using the Chi-square test.
  
  
  RESULTS
  Twenty-one of 31 CD (68%), 13 of 20 UC (65%) and 11 of 23 HC (48%) were MAP-positive (CD vs. HC: p = ns; UC vs. HC: p = ns). With the limits of a small sample size, the IS900-positive percentage in CD and UC was higher than HC, although the difference was not statistically significant.
  
  
  CONCLUSIONS
  The possibility to track the MAP presence in human feces represents a new approach to the "MAP hypothesis". Detection of MAP DNA in feces is very common, reaching very high prevalence both in CD and in UC and even in HC. Our findings seem consistent with a high prevalence of MAP asymptomatic infection among the general population and so the possible involvement of MAP in CD pathogenesis could be linked to a specific immune defective response.
authors.affiliation: Department of Internal Medicine and Gastroenterology, S. Orsola-Malpighi Hospital, Via Massarenti 9, 40138, Bologna, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Tuci
authors.forename: Francesca
authors.initials: F
authors.lastname: Tonon
authors.forename: Lucia
authors.initials: L
authors.lastname: Castellani
authors.forename: Alessandro
authors.initials: A
authors.lastname: Sartini
authors.forename: Giulia
authors.initials: G
authors.lastname: Roda
authors.forename: Margherita
authors.initials: M
authors.lastname: Marocchi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Caponi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Munarini
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Rosati
authors.forename: Giampaolo
authors.initials: G
authors.lastname: Ugolini
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Fuccio
authors.forename: Michele
authors.initials: M
authors.lastname: Scagliarini
authors.forename: Franco
authors.initials: F
authors.lastname: Bazzoli
authors.forename: Andrea
authors.initials: A
authors.lastname: Belluzzi
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001483:Base Sequence / Q000235:genetics*; D016022:Case-Control Studies; D016009:Chi-Square Distribution; D003093:Colitis, Ulcerative / Q000382:microbiology*; D003424:Crohn Disease / Q000382:microbiology*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D003627:Data Interpretation, Statistical; D005243:Feces / Q000737:chemistry / Q000382:microbiology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D016927:Mycobacterium avium subsp. paratuberculosis / Q000235:genetics*; D015203:Reproducibility of Results; D055815:Young Adult
chemical_list: D004269:DNA, Bacterial