Molecular detection of Mycobacterium orygis in lung tissues and lymph nodes using conventional and TaqMan qRT-PCR Assay.

Non-tuberculous mycobacteria detection complicates bovine TB diagnosis in reactor animals.

This study evaluated molecular detection methods for Mycobacterium orygis in bovine tuberculosis (bTB) suspects. Ninety-nine tissue samples (90 lung, 9 lymph node) from cattle and buffaloes were examined using conventional PCR and TaqMan qRT-PCR assays. Following decontamination with 0.75% hexadecylpyridinium chloride and culture on Middlebrook 7H10 media, 26 isolates were recovered. Nineteen isolates tested positive for the hsp65 gene confirming Mycobacterium genus; six were identified as M. orygis and eight as M. kansasii through species-specific PCR. Real-time PCR confirmation of M. orygis isolates yielded cycle threshold values between 18.43-34.74. This research documents the presence of non-tuberculous mycobacteria (NTM) alongside tuberculous mycobacteria in respiratory tissues. The concurrent detection of NTM infections in bTB-reactor animals has diagnostic implications, potentially complicating interpretation of tuberculin skin test results and affecting disease control strategies in cattle and buffalo populations.

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