Evaluation of Surgical Tissue From Patients with Crohnʼs Disease for the Presence of Mycobacterium avium Subspecies paratuberculosis DNA by In Situ Hybridization and Nested Polymerase Chain Reaction

Abstract

Crohn's disease (CD) is a chronic inflammatory bowel disease (IBD) with tissue granuloma and histopathological alteration that resembles aspects in tuberculosis, leprosy, and paratuberculosis. Mycobacterium avium subsp paratuberculosis (MAP) is the causative agent of paratuberculosis, with a suspected role in the etiology of CD. We investigated the presence of MAP DNA in 31 surgical tissue samples from 20 subjects using fluorescence in situ hybridization (FISH) with the aid of confocal scanning laser microscopy and nested polymerase chain reaction (PCR) using the IS900 sequence unique to MAP. MAP DNA was detected by PCR in tissue from 10 of 12 (83%) patients with CD: 7/12 (58%) in inflamed, 6/11 (55%) in noninflamed and in 10 (83%) of either tissue and by FISH in 8 of 12 (67%) patients with CD: 7 of 12 (58%) in inflamed, 4 of 11 (36%) in noninflamed, and in 8(67%) of either tissue. In non-IBD subjects, MAP DNA was detected in the tissue of only 1 of 6 patients (17%) by PCR and 0 of 6 patients (0%) by FISH. MAP DNA was identified by PCR in inflamed tissue from 2 of 2 patients with ulcerative colitis. The detection of MAP DNA by either technique in tissue from subjects with CD is significant compared with non-IBD subjects (P < 0.005). Identification of MAP DNA in both inflamed and noninflamed tissue by both techniques suggests that MAP infection in patients with CD may be systemic. The data add more evidence toward a possible association of MAP in the pathogenesis of CD.

Mycobacterium avium Subspecies paratuberculosis Infection in Cases of Irritable Bowel Syndrome and Comparison with Crohn's Disease and Johne's Disease: Common Neural and Immune Pathogenicities

“…paratuberculosis tests in the gut and the distribution of the gross inflammation in the Crohn's disease patients. This is in close agreement with the results of previous work (55). Mycobacterium avium subsp.…”

Mycobacterium aviumsubsp.paratuberculosisin Lake Catchments, in River Water Abstracted for Domestic Use, and in Effluent from Domestic Sewage Treatment Works: Diverse Opportunities for Environmental Cycling and Human Exposure

“…paratuberculosis is transmitted from time to time to human populations in retail milk supplies (3,24,28,49). Recent research from several centers using appropriate laboratory methods showed that most people with chronic inflammation of the intestine of the Crohn's disease type are infected with this chronic enteric pathogen (2,9,53,63,66,67). Like that of Johne's disease, the incidence of Crohn's disease is increasing (43), particularly in children, with recent data from three sites in Northern and Central Europe and in Australia showing increases in the disease in children under 16 which average fivefold per decade (34,56,59).…”

Search for Localized Dysbiosis in Crohn's Disease Ulcerations by Temporal Temperature Gradient Gel Electrophoresis of 16S rRNA

“…It can be based on culture or on culture-independent methods using specific probes. Despite intensive research focusing on Mycobacteria, Listeria, and Chlamydia, an infectious origin of IBD has not been confirmed (2,13,20,22,30). The global description strategy uses probes for large groups of bacteria which constitute the "dominant microbiota."…”