{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T15:05:13Z","timestamp":1790867113996,"version":"4.1.0"},"reference-count":48,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T00:00:00Z","timestamp":1753660800000},"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:p>\n                    Non tuberculous mycobacteria (NTM) infections are increasing globally, underscoring the critical importance of accurate species-level identification for effective clinical management. This study aimed to evaluate the use of three conserved markers in the mycobacterial family (\n                    <jats:italic>16S<\/jats:italic>\n                    ,\n                    <jats:italic>hsp65<\/jats:italic>\n                    , and\n                    <jats:italic>rpoB<\/jats:italic>\n                    ) for NTM identification through Sanger sequencing, comparing the results to those obtained using MALDI-ToF MS. A total of 59 clinical NTM isolates from plastic surgery patients, previously characterized by MALDI-ToF MS, were analyzed. These isolates underwent DNA extraction, PCR amplification, and Sanger sequencing. Species identification was performed through phylogenetic analyses of each marker individually and concatenated as a multi locus sequencing approach. Concordance between MALDI-ToF MS and Sanger sequencing was assessed using Cohen\u2019s Kappa statistical analysis. Cohen\u2019s Kappa values indicated moderate concordance of 0.46 for\n                    <jats:italic>16S<\/jats:italic>\n                    , 0.51 for\n                    <jats:italic>hsp65<\/jats:italic>\n                    , and 0.69 for\n                    <jats:italic>rpoB<\/jats:italic>\n                    . Concatenated phylogenetic analysis yielded improved concordance values of 0.71 for (\n                    <jats:italic>16S<\/jats:italic>\n                    +\n                    <jats:italic>hsp65)<\/jats:italic>\n                    , 0.76 for (\n                    <jats:italic>16S<\/jats:italic>\n                    +\n                    <jats:italic>rpoB)<\/jats:italic>\n                    , 0.69 for (\n                    <jats:italic>rpoB<\/jats:italic>\n                    +\n                    <jats:italic>hsp65)<\/jats:italic>\n                    , and 0.72 for (16S +\n                    <jats:italic>hsp65<\/jats:italic>\n                    +\n                    <jats:italic>rpoB<\/jats:italic>\n                    ). Our results show that NTM identification is more accurate when employing a multi locus sequencing approach. Notably, the combination of\n                    <jats:italic>16S + rpoB<\/jats:italic>\n                    outperformed the three-marker concatenation, offering the highest concordance for species-level identification. NTM identification is challenging, and concatenated phylogenetic analysis of two or more gene fragments should be used when MALDI-ToF MS or whole genome sequencing is not available.\n                  <\/jats:p>","DOI":"10.3389\/fcimb.2025.1612459","type":"journal-article","created":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T05:30:57Z","timestamp":1753680657000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Comparative evaluation of MALDI-ToF mass spectrometry and Sanger sequencing of the 16S, hsp65, and rpoB genes for non tuberculous mycobacteria species identification"],"prefix":"10.3389","volume":"15","author":[{"given":"Angel Sebastian","family":"Rodriguez-Pazmi\u00f1o","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elsy","family":"Carvajal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Darwin","family":"Paredes-N\u00fa\u00f1ez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose","family":"Echeverr\u00eda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joselyn","family":"Calderon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Solon Alberto","family":"Orlando","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Parra Vera","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miguel Angel","family":"Garcia-Bereguiain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,7,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"5699","DOI":"10.1128\/JCM.41.12.5699-5708.2003\/ASSET\/BE684F4A-DBD4-4FBD-9C88-969A6C9CC686\/ASSETS\/GRAPHIC\/JM1230623003.JPEG","article-title":"rpoB-based identification of nonpigmented and late-pigmenting rapidly growing mycobacteria","volume":"41","author":"Ad\u00e9kambi","year":"2003","journal-title":"J. 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