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Detection of Mycobacteria in Crohn's Disease by a Broad Spectrum Polymerase Chain Reaction

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Background: The role of mycobacterial infection, particularly related to Mycobacterium avium subsp paratuberculosis (Map), in Crohn's disease has long been debated. We developed primer pairs capable of detecting a broad spectrum of mycobacterium and employed them to investigate surgical specimens from patients with Crohn's disease. Methods: Pan mycobacterium primers of the 65-kDa heat shock protein gene (Hsp65) were used in a polymerase chain reaction (PCR) to examine 12 surgically-resected, formalin-fixed, paraffin-embedded specimens from 11 patients with Crohn's disease. The DNA sequences of amplicons were aligned with those in GenBank. Results: Mycobacterial DNA was found in specimens from three of 11 patients. M. mucogenicum was identified in a specimen from one patient and M. tuberculosis in two, but Map was not identified in any. Conclusion: Hsp65-based PCR can be employed to search for occult mycobacterial infection of the gastrointestinal tract in patients with a diagnosis or suspicion of Crohn's disease. This approach may have a therapeutic implication.

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