Development and evaluation of a multiplex molecular point-of-care assay for direct identification ofand prioritized non-tuberculous mycobacteria.

OBJECTIVE: This study aimed to establish a multiplex molecular point-of-care assay calledincorporating an ultra-fast sample pre-treatment for direct identification ofcomplex (MTBC) and 8 non-tuberculous Mycobacteria (NTM) commonly prioritized in clinical settings, and to evaluate its performance in 149 clinical confirmed mycobacterial-positive samples.

METHODS: The study was divided into two stages: a pilot study to establish the methodology and a clinical validation study to evaluate its performance. In the pilot study, we established theand analyzed its performance regarding limits of detection, reproducibility, specificity and efficiency. The clinical validation study was performed using 149 clinical confirmed mycobacterial-positive samples, with 16S rRNA identification as the reference standard. The complete process, from patient to result, was accomplished within 90 minutes.

RESULTS: Of the 149 positive clinical mycobacterial cultures analyzed, 136 were within the designed targets. Among these 136 cultures, 133 samples were correctly identified by, achieving an accuracy rate of 97.79%.

CONCLUSIONS: This study demonstrates thatwith its high accuracy rate are capable to rapidly and effectively differentiate between MTBC and the major NTM species.