Rapid and simultaneous detection of Mycobacterium bovis and Mycobacterium avium subsp. paratuberculosis in invasive flies by duplex PCR.

A rapid and effective polymerase chain reaction (PCR) test, specific for the detection of Mycobacterium bovis and Mycobacterium avium subsp. paratuberculosis in invasive flies, was developed. This duplex PCR technique was specifically designed to amplify the sequences of IS6110 of M. bovis and IS900 of M. avium subsp. paratuberculosis. The results of the interference test, simulated contamination test, and several other experiments indicated that this technique had high stability, specificity, and sensitivity with detection limits at 92.98 pg of M. bovis and 110.27 pg of M. avium subsp. paratuberculosis. Therefore, this duplex PCR test is a rapid and effective technique to simultaneously detect M. bovis and M. avium subsp. paratuberculosis.