Related Experiment Video
Updated: Dec 1, 2025
Identification of Mycobacterium Species by DNA Microarray Chip Method
Published on: June 24, 2025
Identification of nontuberculous mycobacteria species by multiplex real-time PCR with high-resolution melting
Aline Dos Santos Peixoto1,2, Lílian Maria Lapa Montenegro1, Andrea Santos Lima1
1Fundação Oswaldo Cruz, Instituto Aggeu Magalhães, Departamento de Imunologia, Recife, PE, Brasil.
Introduction:
Nontuberculous mycobacteria (NTM) species, as human pathogens, are increasing in the world, as is the difficulty of accurately identifying them. Differential diagnosis, especially between the M. tuberculosis complex and NTM species, and the characterization of NTM species is important. This study aimed to evaluate the performance of a molecular system based on multiplex real-time PCR with high-resolution melting (HRM) for the identification and differentiation of NTM species of clinical importance of an endemic area for tuberculosis in northeastern Brazil.
Methods:
The technical protocol of the molecular system was based on multiplex real-time PCR-HRM, and evaluated the sensitivity and specificity of the detection of NTM species in mycobacterial clinical isolates from the studied region. The gold standard method was specific gene sequencing.
Results:
The sensitivity and specificity of multiplex real-time PCR-HRM modified for differentiation between NTM and M. tuberculosis were 90% and 100%, respectively. The PCR-HRM sensitivities for the characterization of NTM species (M. kansasii, M. abscesses, M. avium, and M. fortuitum) were 94.59%, 80%, 57.14%, and 54%, respectively.
Conclusions:
The multiplex real-time PCR-HRM modified assay has the potential to rapidly and efficiently identify nontuberculous mycobacteria of clinical importance, which is crucial for immediate implementation of the appropriate therapy and thus avoiding complications and sequelae in patients.
