title: Application of multiple laboratory tests for Mycobacterium avium ssp. paratuberculosis detection in Crohn's disease patient specimens.
abstract: The difficulties involved in detecting and enumerating Mycobacterium avium subsp. paratuberculosis (MAP) as a pathogen potentially involved in Crohn's disease (CD) are well known. This study aimed to improve this situation through the application of multiple laboratory diagnostic tests to detect and isolate this bacterium from different specimens collected from CD-patients and non-CD subjects as controls. A total of 120 samples (terminal ileum and colon biopsies, blood and stool) were obtained from 19 CD-patients and from 11 individuals who did not have a clinicopathological diagnosis of CD (non-CD controls) attending for ileocolonoscopy. All samples were processed by staining techniques, culture on both solid and liquid media, and Insertion Sequence 900/F57 real-time PCR. The MAP frequency in CD-patients was found in a significantly greater proportion than in non-CD subjects; the most positive samples were biopsies from CD-patients tested by real-time PCR. MAP detection in biopsies, and in the other samples, by applying multiple and validated laboratory diagnostic tests, could be a marker of active infection, supporting MAP involvement in CD.
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Banche
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Allizond
authors.affiliation: Gastroenterology Division, AO Ordine Mauriziano, Turin, Italy.
authors.forename: Raffaello
authors.initials: R
authors.lastname: Sostegni
authors.affiliation: Gastroenterology Division, AO Ordine Mauriziano, Turin, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Lavagna
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Bergallo
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Sidoti
authors.affiliation: Gastroenterology Division, AO Ordine Mauriziano, Turin, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Daperno
authors.affiliation: Gastroenterology Division, AO Ordine Mauriziano, Turin, Italy.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Rocca
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy.
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Cuffini
mesh_terms: D000328:Adult; D000368:Aged; D003424:Crohn Disease / Q000175:diagnosis / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D016927:Mycobacterium avium subsp. paratuberculosis / Q000235:genetics / Q000302:isolation & purification*; D010283:Paratuberculosis / Q000175:diagnosis / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction; D055815:Young Adult
chemical_list: D004269:DNA, Bacterial
keywords: Biopsies; Crohn’s disease; Culture; IS900/F57; Mycobacterium avium subsp. paratuberculosis; Real-time PCR; Staining techniques; Stool