Differential Identification of Mycobacterial Species Using High-Resolution Melting Analysis.

Infections caused by non-tuberculous mycobacteria (NTM) is increasing wordwide. Due to the difference in treatment of NTM infections and tuberculosis, rapid species identification of mycobacterial clinical isolates is necessary for the effective management of mycobacterial diseases treatment and their control strategy. In this study, a cost-effective technique, real-time PCR coupled with high-resolution melting (HRM) analysis, was developed for the differentiation of Mycobacterial species using a novel sequence. A total of 107 mycobacterial isolates (nine references and 98 clinical isolates) were subjected to differentiation using locus sequence in a real-time PCR-HRM assay scheme. From 98 Mycobacterium clinical isolates, 88 species (89.7%), were identified at the species level by locus sequence analysis as a gold standard method. was the most frequently encountered species (41 isolates), followed by (20 isolates), (15 isolates), (10 isolates), group (5 isolates), (5 isolates), and and one isolate each. The HRM analysis generated six unique specific groups representing complex, , , , - group, and complex. In conclusion, this study showed that the -based real-time PCR followed by HRM analysis could differentiate the majority of mycobacterial species that are commonly encountered in clinical specimens.