Abstract

The association of Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) with Crohn’s disease is a controversial issue. M. paratuberculosis is detected by amplifying the IS900 gene, as microbial culture is unreliable from humans. We determined the presence of M. paratuberculosis in patients with Crohn’s disease (CD) (n = 22), ulcerative colitis (UC) (n = 20), aphthous ulcers (n = 21) and controls (n = 42) using PCR assays validated on bovine tissue. Culture from human tissue was also performed. M. paratuberculosis prevalence in the CD and UC groups was compared to the prevalence in age and sex matched non-inflammatory bowel disease controls. Patients and controls were determined to be M. paratuberculosis positive if all three PCR assays were positive. A significant association was found between M. paratuberculosis and Crohn’s disease (p = 0.02) that was not related to age, gender, place of birth, smoking or alcohol intake. No significant association was detected between M. paratuberculosis and UC or aphthous ulcers; however, one M. paratuberculosis isolate was successfully cultured from a patient with UC. We report the resistance of this isolate to ethambutol, rifampin, clofazamine and streptomycin. Interestingly this isolate could not only survive but could grow slowly at 5°C. We demonstrate a significant association between M. paratuberculosis and CD using multiple pre-validated PCR assays and that M. paratuberculosis can be isolated from patients with UC.

Highlights

  • Inflammatory Bowel Disease (IBD) and its divisions of Crohn’s disease (CD) and ulcerative colitis (UC) often strike in the prime of life and remain a life-long burden [1]

  • A number of microorganisms, including Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis), have been associated with IBD, but as yet, evidence to support the role of a specific microorganism in IBD is missing

  • For the aphthous ulcers group, 2/21 patients were positive with the single round IS900 only and no other PCR assay, this group was not compared to a control group

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Summary

Introduction

Inflammatory Bowel Disease (IBD) and its divisions of Crohn’s disease (CD) and ulcerative colitis (UC) often strike in the prime of life and remain a life-long burden [1]. While the aetiology of IBD remains unclear, there is strong evidence to support the role of both microorganisms [2] and host genetic factors [3,4,5,6]. Paratuberculosis (M. paratuberculosis), have been associated with IBD, but as yet, evidence to support the role of a specific microorganism in IBD is missing. The prevalence of M. paratuberculosis in patients with CD and UC has been shown to be highly variable (92% and 0–35%, respectively) [7,8,9], a meta-analysis in 2007 [10] demonstrated a significant association between M. paratuberculosis and CD. The majority of studies investigating the association of M. paratuberculosis and CD have used one marker, IS900, a unique insertion sequence of M. paratuberculosis [15,16,17]. A pure isolate of M. paratuberculosis was obtained from a patient with UC and a comprehensive biochemical and molecular characterisation of the second subculture of this isolate is reported

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