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Absence of Mycobacterium avium subspecies paratuberculosis Components from Crohn’s Disease Intestinal Biopsy Tissues

Department of Veterinary Pathology, College of Veterinary Medicine, Iowa State University, Ames, Iowa and the Center for Veterinary Biologics Laboratory, Veterinary Services, Animal and Plant Health Inspection Service, United States Department of Agriculture, Ames, Iowa

John C. Cheville, MD

Mayo Clinic, Rochester, Minnesota

Dominique Brees, DVM, PhD. DACVP

Department of Veterinary Pathology, College of Veterinary Medicine, Iowa State University, Ames, Iowa

Janice M. Miller, DVM, PhD

Zoonotic Diseases Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Ames, Iowa

Norman F. Cheville, DVM, PhD, DrHc

Department of Veterinary Pathology, College of Veterinary Medicine, Iowa State University, Ames, Iowa

REPRINT REQUESTS: Jay L.E. Ellingson, PhD, Marshfield Clinic Laboratories, Food Safety Services, Marshfield Clinic, 1000 North Oak Avenue, Marshfield, WI 54449, Telephone: 715-389-5958, Fax: 715-389-7599, Email: ellingson.jay{at}marshfieldclinic.org

BACKGROUND

Crohn’s disease is a chronic human intestinal inflammatory disorder for which an etiologic agent has not been identified. Johne’s disease is a similar chronic enteric granulomatous disease of ruminant species and has been used as a model of Crohn’s disease. Johne’s disease has been proven to be caused by Mycobacterium avium subspecies paratuberculosis (M. avium ss paratuberculosis). It has been proposed that M. avium ss paratuberculosis may also cause Crohn’s disease. This is of particular concern because the organism may be spread to humans through inadequately pasteurized dairy products.

OBJECTIVE

We sought to determine whether M. avium ss paratuberculosis could be detected using identical techniques in paraffin-embedded tissue samples of bovine Johne’s disease and human Crohn’s, ulcerative colitis and diverticular diseases. Samples were obtained for analysis from national tissue banks.

DESIGN

Cross-species and cross-disease sample comparisons by multiple detection techniques.

METHODS

Histology, immunocytochemistry and polymerase chain reaction (PCR) were utilized to test and compare the presence of M. avium ss paratuberculosis components. Insertion sequence IS900, present in multiple copies and found only in M. avium ss paratuberculosis, was utilized in both PCR and immunocytochemical analyses.

RESULTS

The IS900 sequence was demonstrable in all samples of confirmed positive Johne’s disease tissue. The sequence was not identified in the 35 Crohn’s, 36 ulcerative colitis, and 21 diverticular disease samples.

CONCLUSION

M. avium ss paratuberculosis was not associated with the lesions in these Crohn’s disease samples, using these methods.

Key Words: Crohn’s disease • Johne’s disease • Inflammatory bowel disease • Mycobacterium avium subspecies paratuberculosis • IS900 • Polymerase chain reaction