Prevalence of Mycobacterium avium subspecies paratuberculosis IS 900 DNA in biopsy tissues from patients with Crohn’s disease: histopathological and molecular comparison with Johne’s disease in Fars province of Iran

Abstract

Abstract Background Crohn’s disease is a chronic enteritis of humans that affects the gastrointestinal tract, especially the terminal ileum, cecum and colon. The etiology of this disease is still unknown but seems to be multifactorial. There are reports about the potential link between Crohn’s disease in humans and the causative agent of Johne’s disease in ruminants. Because of the prevalence of Johne’s disease in the Fars Province of Iran, the aim of this study was to investigate the prevalence of MAP in the biopsy tissues of patients affected by Crohn’s disease in this area. Methods The study was performed from April 2015 to June 2017 at Namazi Hospital, Shiraz University of Medical Sciences, and School of Veterinary Medicine, Shiraz University, Shiraz, Iran. Intestinal biopsies of 30 patients (12 male and 18 female; mean age, 34 years; range 4–77 years) with the confirmed diagnosis of Crohn’s disease and 30 patients diagnosed as non-inflammatory bowel disease (19 male and 11 female; mean age, 38 years; range 13–68 years) were studied by molecular, histopathological and histochemical methods. Also, similar numbers of adult goats affected by Johne’s disease were studied, comparatively. DNA extractions of tissue specimens were subjected to PCR to amplify a 413-bp sequence of the IS900 gene. Results Using IS900-PCR, the overall prevalence of MAP in patients affected by Crohn’s disease and non-inflammatory bowel disease were 47 and 13%, respectively. In addition, the prevalence of MAP in goats affected by Johne’s disease was 70%. Using acid-fast histochemical staining, only 7% of Crohn’s disease patients were weakly positive as paucibacillary and 43% of Johne’s disease cases were moderate to strongly positive as multibacillary. Histopathologically, granulomatous enteritis (83 and 90%), lymphoplasmacytic enteritis (17 and 14%), edema and lymphangiectasia (67 and 96%), and vasculitis (20 and 73%) were common findings in Crohn’s and Johne’s diseases, respectively. Conclusion Our findings demonstrate a remarkable association between MAP and CD in this population, and support an etiologic relationship between MAP infection in humans and the development of CD. MAP infection in human tissue may display species-specific pathologic findings, as occurs with other zoonotic pathogens.

Keywords

• IS900-PCR
• Crohn’s disease
• Johne’s disease
• Mycobacterium avium subsp. paratuberculosis