Analysis of three cases with false positive PCR results of non tuberculosis mycobacterium

Real-time fluorescent quantitative PCR (RT-PCR) can effectively distinguish between Mycobacterium tuberculosis (MTB) and Non-tuberculosis mycobacterium (NTM), but when there are overlapping sequences between other pathogens (such as Nocardia otidiscaviarum, Mycobacterium parantracellulare, Mycolicibacterium fluoranthenivorans) and NTM, abnormal amplification curves may appear.

The clinical manifestations of the three patients were fever and respiratory symptoms. Chest CT showed "multiple lung infections". The acid-fast bacilli were negative by microscopic examination. The results of RT-PCR detection of Mycobacterium tuberculosis DNA showed that they are all NTM, while the results of DNA microarray method showed that there were no non-Mycobacterium tuberculosis. Identified by MALDI-TOF mass spectrometry, they are Nocardia otidiscaviarum, Mycobacterium parantracellale, Mycolicibacterium fluoranthenivorans. We found that the sequences of the above three bacteria can be combined with the primers and probes used for NTM PCR detection, resulting in false positive.

In the RT-PCR detection of mycobacteria, if there's abnormal amplification, and the mycobacterial species cannot be identified, the amplified products sequencing or MALDI- TOF mass spectrometry identification will help avoid the omission of rare pathogens.