Rapid detection of rifampin resistance in Mycobacterium tuberculosis using nucleotide MALDI-TOF MS: a comparative study with phenotypic drug susceptibility testing and DNA sequencing

. Its high concordance with DNA sequencing, excellent diagnostic performance, and ability to identify heteroresistance highlight its potential as a valuable tool for early TB diagnostics and improve the precision of chemotherapy regimen development.IMPORTANCEThe emergence of multidrug-resistant tuberculosis (MDR-TB) and rifampin-resistant tuberculosis (RR-TB) poses a significant challenge to global tuberculosis (TB) control efforts. Rifampin (RIF) resistance is a critical marker for MDR-TB, which requires more complex, prolonged, and costly treatment regimens. Early and accurate detection of RIF resistance is crucial for effective TB control. This study evaluates the performance of nucleotide MALDI-TOF MS, an innovative technology, for detecting RIF resistance-associated mutations in the rpoB gene. The method demonstrates high sensitivity (93.2%) and specificity (98.1%), with the added advantage of identifying heteroresistance, capabilities that are lacking in conventional methods. These capabilities are crucial for early diagnosis, guiding personalized treatment regimens, and curbing the transmission of drug-resistant TB. The findings demonstrate that nucleotide MALDI-TOF MS provides a rapid, high-throughput, and cost-effective alternative for detecting rpoB gene mutations associated with RIF resistance.