A novel one-tube nested fluorescence melting curve analysis for rapid detection of drug-resistant Mycobacterium tuberculosis.

ABSTRACT:

Background

Rapidly detecting drug-resistant Mycobacterium tuberculosis (MTB) is essential for effective tuberculosis (TB) treatment. We have developed a one-tube nested fluorescence melting curve assay (OTN-FMCA) that allows rapid and simultaneous detection of resistance to rifampicin, isoniazid, fluoroquinolones, and bedaquiline.

Results

The OTN-FMCA uses nested PCR and locked nucleic acid (LNA)-modified probes to identify 44 mutations in six key genes (rpoB, katG, inhA, gyrA, gyrB, and atpE), achieving a detection limit of 5 copies per microliter and identifying up to 10% heterogeneous resistance. When tested on 198 culture-positive isolates, OTN-FMCA demonstrated high concordance with microarray susceptibility testing, with 100% sensitivity and 96.89% specificity. Further

SUBMITTER: Chang Y 

PROVIDER: S-EPMC12859952 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature