{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T23:45:25Z","timestamp":1782949525534,"version":"3.54.5"},"reference-count":69,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,3,25]],"date-time":"2026-03-25T00:00:00Z","timestamp":1774396800000},"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. Tuberc."],"abstract":"<jats:p>\n                    Nontuberculous mycobacteria (NTM) represent an increasingly significant cause of pulmonary and extrapulmonary infections, but are sometimes misinterpreted as tuberculosis (TB) owing to overlapping clinical and microbiological characteristics. Conventional diagnostic approaches, such as Ziehl-Neelsen staining and culture in a Mycobacterial Growth Indicator Tube (MGIT) system, are constrained by extended incubation times, are insufficient for accurate species differentiation, and are limited by prolonged incubation periods. Recent molecular and genomic advances have transformed NTM diagnostics by enabling rapid, specific, and high-resolution identification. Line probe assays (e.g., GenoType Mycobacterium CM\/AS assay) and multiplex PCR have enhanced the ability to distinguish between NTM species such as\n                    <jats:italic>Mycobacterium absessus, M. fortuitum<\/jats:italic>\n                    , and\n                    <jats:italic>M. avium<\/jats:italic>\n                    complex and\n                    <jats:italic>M. tuberculosis<\/jats:italic>\n                    complex, which is essential for proper treatment and epidemiological mapping. Among newer proteomic platforms, matrix-assisted laser desorption\/ionization time-of-flight (MALDI-TOF) mass spectrometry has emerged as a transformative, cost-effective technology capable of identifying\n                    <jats:italic>Mycobacterium<\/jats:italic>\n                    species directly from culture isolates through protein fingerprinting. It provides rapid, reproducible, and highly discriminatory identification between closely related species. Next-generation sequencing (NGS) and whole genome sequencing approaches now offer unprecedented insight into species identification, strain typing, and drug-resistance prediction, complementing traditional culture-based susceptibility testing. Newer techniques such as metagenomics NGS (mNGS), targeted NGS (tNGS) multilocus sequence typing, and mycobacterial interspersed repetitive unit-variable number tandem repeats\n                    <jats:bold>(<\/jats:bold>\n                    MIRU-VNTR) genotyping facilitate subspecies-level resolution and real-time outbreak surveillance. Moreover, molecular beacons, insertion sequence analysis, and repetitive sequence-based polymerase chain reaction (Rep-PCR) enhance detection sensitivity even in paucibacillary samples. The integration of genomic data with automated diagnostic system promises earlier intervention, accurate species delineation, and improved patient outcome.\n                  <\/jats:p>","DOI":"10.3389\/ftubr.2026.1760581","type":"journal-article","created":{"date-parts":[[2026,3,25]],"date-time":"2026-03-25T06:55:08Z","timestamp":1774421708000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Advanced diagnostic methods for nontuberculous mycobacterial infections"],"prefix":"10.3389","volume":"4","author":[{"given":"Sneha","family":"Singh","sequence":"first","affiliation":[{"name":"Departments of Zoology and Environmental Sciences, Deen Dayal Upadhyaya Gorakhpur University","place":["Gorakhpur, Uttar Pradesh, India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amresh Kumar","family":"Singh","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Baba Raghav Das Medical College","place":["Gorakhpur, Uttar Pradesh, India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sushil","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Zoology, Deen Dayal Upadhyaya Gorakhpur University","place":["Gorakhpur, Uttar Pradesh, India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nandini","family":"Singh","sequence":"additional","affiliation":[{"name":"Department of Zoology, Deen Dayal Upadhyaya Gorakhpur University","place":["Gorakhpur, Uttar Pradesh, India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashwini Kumar","family":"Mishra","sequence":"additional","affiliation":[{"name":"Department of Tuberculosis & Chest, Baba Raghav Das Medical College","place":["Gorakhpur, Uttar Pradesh, India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aroop","family":"Mohanty","sequence":"additional","affiliation":[{"name":"Department of Microbiology, All India Institute of Medical Sciences","place":["Gorakhpur, Uttar Pradesh, India"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,3,25]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/biology10020096","article-title":"Drug resistance in nontuberculous mycobacteria: mechanisms and models","volume":"10","author":"Saxena","year":"2021","journal-title":"Biology."},{"key":"B2","doi-asserted-by":"publisher","first-page":"949","DOI":"10.1080\/17476348.2021.1925545","article-title":"Non-tuberculous mycobacteria: a disease beyond TB and preparedness in India","volume":"15","author":"Sharma","year":"2021","journal-title":"Expert Rev Respir Med."},{"key":"B3","doi-asserted-by":"publisher","first-page":"083","DOI":"10.4103\/0974-2727.119847","article-title":"Role of GenoType\u00ae mycobacterium common mycobacteria\/additional species assay for rapid differentiation between Mycobacterium tuberculosis complex and different species of non-tuberculous mycobacteria","volume":"5","author":"Singh","year":"2013","journal-title":"J Lab Physicians."},{"key":"B4","doi-asserted-by":"publisher","first-page":"e81017","DOI":"10.7759\/cureus.81017","article-title":"Extrapulmonary nontuberculous mycobacteria infection: the new-age neglected infectious disease","volume":"17","author":"Rath","year":"2025","journal-title":"Cureus"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1183\/23120541.00336-2022","article-title":"Global burden of nontuberculous mycobacteria in the cystic fibrosis population: a systematic review and meta-analysis","author":"Prieto","year":"2023","journal-title":"ERJ Open Res"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ccm.2014.10.002","article-title":"Marras TK. 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