We have developed a rapid diagnostic method, based on a real-time PCR and melting temperature analysis of amplicons, for detecting Mycobacterium avium subspecies paratuberculosis (Map) DNA in faecal samples. The primers to amplify the IS900 gene segment have shown a high sensitivity of 0.001-0.0005 pg Map DNA/PCR and a specificity with no cross-reactions among 58 mycobacterial strains tested. As compared with the conventional bacterial culture method, which requires more than two months to obtain a result, this detection method using a real-time PCR can diagnose Johne's disease within several hours.
(Research Team for Paratuberculosis, TEL +81-29-838-7857)