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Whole-genome sequencing (WGS) was applied to assess phylogenetic relatedness, identify virulence factors, and characterize the resistome of the isolates. Antimicrobial susceptibility testing (AST) was performed using the GenoType NTM-DR line probe assay (LPA), Sensititre minimum inhibitory concentrations (MIC) plates, and the proportional method based on critical concentrations. Spatial distribution of cases was mapped using ArcGIS software.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Spatiotemporal distribution patterns indicated the presence of circulating clones confined within specific geographical areas. Plasmids coding for ferredoxin and cytochrome P450 genes were identified in one cluster, which notably lacked the chromosomal\n                      <jats:italic>mbtH<\/jats:italic>\n                      gene involved in siderophore biosynthesis for iron acquisition. In contrast, isolates grouped in a second cluster harbored the\n                      <jats:italic>mbtH<\/jats:italic>\n                      chromosomal gene but lacked these plasmid-associated elements. LPA and broth microdilution showed that all\n                      <jats:italic>Mycobacterium monacense<\/jats:italic>\n                      isolates were susceptible to aminoglycosides, fluoroquinolones, and macrolides, but generally exhibited elevated MICs against \u03b2-lactam antibiotics. Phenotypic AST indicated that drugs commonly used to treat\n                      <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                      complex (MTBC), namely bedaquiline, linezolid, and rifampicin, are effective against\n                      <jats:italic>Mycobacterium monacense<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Mycobacterium monacense<\/jats:italic>\n                      in extrapulmonary cultures accentuates the need for improved diagnostics and enhanced clinical awareness of infections with rare NTM. WGS highlights the potential significance provided by plasmid-encoded genes. Current treatment regimens for MTBC exhibit therapeutic efficacy against\n                      <jats:italic>Mycobacterium monacense<\/jats:italic>\n                      isolates.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s10096-025-05318-y","type":"journal-article","created":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T12:37:38Z","timestamp":1761655058000},"page":"403-417","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Clonal spread of Mycobacterium monacense strains among patients with extrapulmonary mycobacteriosis"],"prefix":"10.1007","volume":"45","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6676-5431","authenticated-orcid":false,"given":"Christoffel","family":"Opperman","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-1014-5104","authenticated-orcid":false,"given":"Aysha","family":"Ahmed","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-4708-4537","authenticated-orcid":false,"given":"Marianna","family":"De Kock","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9580-8777","authenticated-orcid":false,"given":"Claudia","family":"Spies","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1337-2132","authenticated-orcid":false,"given":"Michael","family":"Biggel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8005-062X","authenticated-orcid":false,"given":"Tristen","family":"Lourens","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1794-4126","authenticated-orcid":false,"given":"Sarishna","family":"Singh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2002-5240","authenticated-orcid":false,"given":"Yonas","family":"Ghebrekristos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5741-7358","authenticated-orcid":false,"given":"Robin","family":"Warren","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6614-9084","authenticated-orcid":false,"given":"Wynand","family":"Goosen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3936-9687","authenticated-orcid":false,"given":"Giovanni","family":"Ghielmetti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,10,28]]},"reference":[{"issue":"3","key":"5318_CR1","doi-asserted-by":"publisher","first-page":"475","DOI":"10.3390\/microorganisms13030475","volume":"13","author":"M Ramesh","year":"2025","unstructured":"Ramesh M, Behra PRK, Pettersson BMF, Dasgupta S, Kirsebom LA (2025) Age-dependent pleomorphism in Mycobacterium monacense cultures. 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