Triplex PCR for Detection of Mycobacterium spp. in Blood of Bovine TB Reactors

The presence of M. tuberculosis in bovines indicates the process of reverse zoonosis as animals infected can transmit it back to humans. Therefore, early, fast and precise diagnosis becomes an important step. The present study was conducted to detect Mycobacteria in blood samples of bTB reactors in which a total of 37 samples were tested using Triplex PCR by targeting the internal transcribed spacer (ITS) region between the 16S and 23S r-DNA genes. It was followed by nested PCR by using three specific amplifications targeting Mycobacteria, M. bovis/M. tuberculosis and MAC with base pair sizes of 322bp, 133bp and 83bp respectively. Out of 37 blood samples, 12 and 5 samples were found exclusively positive for M. bovis/M. tuberculosis and MAC respectively, whereas one sample was positive for both. Triplex PCR was found to be useful for the detection of co-infections of mycobacterial infections in blood samples of bTB reactor animals.