Rapid Identification of Clinically Relevant Mycobacterium Species by Multicolor Melting Curve Analysis.

The sustained increase in nontuberculous mycobacteria (NTM) infection incidence and the difficulty in distinguishing these infections from tuberculosis constitute an urgent need for NTM species level identification. The MeltPro Myco assay is the first diagnostic system that identifies 19 clinically relevant mycobacteria in a single reaction based on multicolor melting curve analysis run on a real-time PCR platform. The assay was comprehensively evaluated regarding its analytical and clinical performances. The MeltPro Myco assay accurately identified 51 reference mycobacterial strains to the species/genus level and showed no cross-reactivity with 16 non-mycobacterial strains. The limit of detection was 300 bacilli/mL and 1% of the minor species was detected in the case of mixed infections. Clinical studies using 1,163 isolates collected from five geographically distinct healthcare units showed that the MeltPro Myco assay correctly identified 1,159 (99.7%) samples. Further testing with 94 smear-positive sputum samples showed that all samples were correctly identified. Additionally, the entire assay can be performed within 3 h. The results of this study confirmed the efficacy of this assay in reliable identification of mycobacteria, suggesting that it might potentially be used as a screening tool in tuberculosis-endemic regions.