High Sensitivity of Targeted Next-Generation Sequencing on Patients with Suspected Mycobacteria Infectious Diseases: A Prospective Matched Cohort Study

Objectives To evaluate the diagnostic accuracy of Mycobacterium -targeted next-generation sequencing (M-tNGS)technique forpatients with suspected Mycobacterium infectious diseases. Methods We consecutively enrolled patients with suspected Mycobacterium infectious diseases, collected samples (respiratory tract sample and tissue biopsy), and tested them using M-tNGS and GeneXpert MTB RIF (Xpert) assays. The results of these tests were compared with the clinical diagnosis and Mycobacterium culture results. Results Overall,244 patients were enrolled, from whom 206 respiratory samples and 38 tissue samples were obtained. The sensitivity of M-tNGS was superior to that of Xpert in both respiratory and biopsy samples (92.2% vs. 43.6% and 90.0% vs. 46.0%, respectively). Conversely, M-tNGS was less specific than Xpert in respiratory and biopsy samples (79.7% vs. 100% and 87.5% vs. 100% , respectively). Meanwhile, M-tNGS more frequently detected drug resistance and nontubercular mycobacteria (NTM), with sensitivity of 70.91% and 13.11%, respectively. Based on comparison with a composite reference standard, M-tNGS was more accurate than the Xpert assay and Mycobacteriumtuberculosis(MTB) culture, with areas under the curve of 0.86, 0.72, and 0.6, respectively. Conclusion M-tNGS is suitable for the early screening of Mycobacterium infectious diseases. Notably, M-tNGS can provide more information on drug resistance and Mycobacterium species identification, facilitating accurate treatment.