{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T15:41:58Z","timestamp":1790178118342,"version":"4.1.0"},"reference-count":35,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,8,13]],"date-time":"2025-08-13T00:00:00Z","timestamp":1755043200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>This study aimed to establish a multiplex molecular point-of-care assay called <jats:italic>fastNTM<\/jats:italic> incorporating an ultra-fast sample pre-treatment for direct identification of <jats:italic>Mycobacterium tuberculosis<\/jats:italic> complex (MTBC) and 8 non-tuberculous Mycobacteria (NTM) commonly prioritized in clinical settings, and to evaluate its performance in 149 clinical confirmed mycobacterial-positive samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The study was divided into two stages: a pilot study to establish the methodology and a clinical validation study to evaluate its performance. In the pilot study, we established the <jats:italic>fastNTM<\/jats:italic> and analyzed its performance regarding limits of detection, reproducibility, specificity and efficiency. The clinical validation study was performed using 149 clinical confirmed mycobacterial-positive samples, with 16S rRNA identification as the reference standard. The complete process, from patient to result, was accomplished within 90 minutes.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Of the 149 positive clinical mycobacterial cultures analyzed, 136 were within the designed targets. Among these 136 cultures, 133 samples were correctly identified by <jats:italic>fastNTM<\/jats:italic>, achieving an accuracy rate of 97.79%.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>This study demonstrates that <jats:italic>fastNTM<\/jats:italic> with its high accuracy rate are capable to rapidly and effectively differentiate between MTBC and the major NTM species.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2025.1560870","type":"journal-article","created":{"date-parts":[[2025,8,13]],"date-time":"2025-08-13T05:33:40Z","timestamp":1755063220000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Development and evaluation of a multiplex molecular point-of-care assay for direct identification of Mycobacterium tuberculosis and prioritized non-tuberculous mycobacteria"],"prefix":"10.3389","volume":"15","author":[{"given":"Qiao-Lian","family":"Yi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yun","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuang","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng-Li","family":"Feng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao-Yu","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin-Zhu","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao-Tian","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Fan","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi-Wen","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying-Chun","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,8,13]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"2095","DOI":"10.1099\/ijs.0.63094-0","article-title":"Dissection of phylogenetic relationships among 19 rapidly growing Mycobacterium species by 16S rRNA, hsp65, sodA, recA and rpoB gene sequencing","volume":"54","author":"Ad\u00e9kambi","year":"2004","journal-title":"Int. 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