Drug resistance of Mycobacterium tuberculosis in West Java, Indonesia

Tuberculosis (TB) is currently one of the leading causes of infectious disease deaths globally, and Indonesia ranks 2nd in annual TB cases, below only India. Accurate TB diagnosis and detection of multidrug-resistant TB (MDR-TB) in real-world settings are crucial for prompt treatment and surveillance. We therefore compared multiple methods for TB detection and drug resistance profiling, including a cartridge-based nucleic acid amplification test (CBNAAT), line probe assay (LPA), and phenotypic drug susceptibility testing (pDST) with targeted long-read next generation sequencing (tNGS) and whole genome sequencing (WGS) on 133 patients in West Java, Indonesia. WGS enabled comprehensive phylogenetic analyses and insights into TB evolution and drug resistance patterns, but its low read counts limit practicality for clinical use. Comparatively, tNGS demonstrated superior sensitivity and specificity, effectively identifying resistance profiles across multiple first-line and second-line drugs with rapid turnaround times. Notably, when compared to LPA, tNGS showed positive percent agreement (PPA) values of 100 % for rifampicin, isoniazid and ethionamide, and an overall agreement of 94 % across multiple drugs. In comparison with CBNAAT, the tNGS PPA for rifampicin remained high at 91 %. The results show that long-read tNGS technology offers a robust tool for enhanced TB treatment and surveillance, ensuring both timely detection and enabling effective tracing through in-depth genetic analysis. The findings significantly contribute to the development of strategies for TB control and management, especially in regions with a high burden of TB cases.