Specific detection of Mycobacterium paratuberculosis DNA associated with granulomatous tissue in Crohn's disease.

Fidler, Helen; Thurrell, Wendy; Johnson, N M; Rook, G.A.W.; McFadden, Johnjoe

doi:10.1136/gut.35.4.506

Cited by 101 publications

(59 citation statements)

References 21 publications

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“…14 The majority of evidence is based on detection of the organism in Crohn's disease patients by immunologic and molecular methods. [15][16][17][18] For example, the IS900 insertion sequence has been detected by polymerase chain reaction (PCR) in intestinal biopsy specimens of Crohn's disease patients. 14,16 However, many of these studies are conflicting and utilize a variety of laboratory techniques without culture confirmation.…”

Section: Introductionmentioning

confidence: 99%

Growth, Congo Red Agar Colony Morphotypes and Antibiotic Susceptibility Testing of Mycobacterium avium subspecies paratuberculosis

Parrish¹,

Ko²,

Dick³

et al. 2004

Clinical Medicine & Research

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OBJECTIVEMycobacterium avium subspecies (subsp.) paratuberculosis (MAP) is the causative agent of Johne's disease in ruminants and has been associated with Crohn's disease in humans. We sought to test growth rates and susceptibilities of various strains of MAP in two available growth media. METHODSUsing the BACTEC macrobroth radiometric growth system and Congo Red-staining agar media, we determined inherent differences in growth characteristics of three bovine and two human strains of MAP and compared susceptibility results obtained in each growth system. RESULTSSignificant differences were observed in growth rate as well as mycobactin J dependence between strains and between a laboratory-adapted isolate of the same strain in the macrobroth system. Similarly, colonial morphology and Congo Red staining on agar media were observed. Two strains, one human and one bovine, demonstrated a 100% rough transparent colony with white coloration on Congo Red agar, while one bovine isolate exclusively grew as a smooth opaque colony with red coloration on Congo Red agar. The remaining strains exhibited mixtures of these two colonial morphotypes on agar media. Comparative susceptibility results between the BACTEC radiometric macrobroth method and the agar proportionality method showed good correlation for most antibiotics/inhibitors tested. However, erratic or poor growth in the macrobroth system prevented minimal inhibitory concentration determinations for two bovine strains by this method. CONCLUSIONThis study demonstrates the variability in the colonial morphology of MAP on Congo Red agar as well as the correlation of antibiotic susceptibility results between the BACTEC macro broth method and the agar proportionality method. This study also emphasizes the need for the development of improved, standardized culture and susceptibility test methods for MAP.

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Section: Introductionmentioning

confidence: 99%

Growth, Congo Red Agar Colony Morphotypes and Antibiotic Susceptibility Testing of Mycobacterium avium subspecies paratuberculosis

Parrish¹,

Ko²,

Dick³

et al. 2004

Clinical Medicine & Research

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“…paratubercuhsis from patients with CD [10] sparked renewed interest in this organism in relation to CD. However, detection of mycobacterial DNA in CD tissue by amplification using Ihe polymerase chain reaction (PCR) has produced conflicting results [11][12][13][14][15][16][17].…”

Section: Introductionmentioning

confidence: 99%

Peripheral cell-mediated immune response to mycobacterial antigens in inflammatory bowel disease

Rowbotham

Howdle

Trejdosiewicz

1995

Clinical and Experimental Immunology

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SUMMARYA mycobacterial etiology has been proposed In Crohn's disease (CD). We have sought evidence of increased or modified T lymphocyte immune responses to Mycnhactcrium tuberculosis and Myco. paratubercuhsis in patients with CD (ti = 13), compared with ulcerative colitis (UC; n = 17) and controls (« = 17). Peripheral blood cells were cultured with phytohaemagglutinin (positive mitogen control), mycobacterial purified protein derivative (PPD) preparations, lysates., column fractions and whole, heat-killed bacteria. Responses of T cells and T cell subsets were assessed by expression of activation markers (CD25. CD69). coupled with blast ogenesis assays (^H-thymidine uptake) and estimates of proliferation. Virtually all patients responded to Myco. paratuberculo.\is and Myco. tuberculosis antigens. There were no significant differences between patient groups, although there was a very high overall correlation (r = 095; P<0000\) between responses to the two mycobacterial species. Most of the activation and proliferative responses resided in the CD4'*' (T helper) subset. Although up to 15% of CDS"^ (suppressor/cytotoxic) cells also became activated, the CD8^ cells did not proliferate subsequently. Cells expressing the alternate y/) form of the T cell receptor (TCR 7(S^)did not activate or proliferate in response to mycobacterial antigens. There were no differences in any of these parameters between patient groups. We conclude that there is no specilic increase or alteration in cell-mediated anti-mycobacterial immunity in inflammatory bowel disease (IBD). Thus our data do not suppwrt a mycobacterial etiopathology of Crohn's disease.

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“…4,6,7,9,15,16,19 Other authors have found a variable number of cases, up to 100%, to harbor the organism. 1,3,5,8,[10][11][12][13][14]18 Several reasons may be responsible for the variability in these results. Differences in patient cohorts, techniques, and experimental conditions immediately come to mind.…”

Section: Discussionmentioning

confidence: 99%

“…3,7,8,[10][11][12][13][14] Thus, the finding of MAP DNA in Crohn's disease, including in paraffin-embedded resection specimens, 1,12 still leaves unanswered the question of whether the organism may be environmentally acquired and nonspecifically present. MAP has been cultured from municipal water supplies 11 and pasteurized milk, 25,26 suggesting that it may be ubiquitously present in the environment.…”

Section: Discussionmentioning

confidence: 99%

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Absence of Mycobacterium avium subsp. paratuberculosis in the microdissected granulomas of Crohn's disease

et al. 2004

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Specific detection of Mycobacterium paratuberculosis DNA associated with granulomatous tissue in Crohn's disease.

Abstract: The role of mycobacteria,

Cited by 101 publications

References 21 publications

Growth, Congo Red Agar Colony Morphotypes and Antibiotic Susceptibility Testing of Mycobacterium avium subspecies paratuberculosis

Growth, Congo Red Agar Colony Morphotypes and Antibiotic Susceptibility Testing of Mycobacterium avium subspecies paratuberculosis

Peripheral cell-mediated immune response to mycobacterial antigens in inflammatory bowel disease

Absence of Mycobacterium avium subsp. paratuberculosis in the microdissected granulomas of Crohn's disease

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