{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,7]],"date-time":"2026-05-07T14:01:12Z","timestamp":1778162472992,"version":"3.51.4"},"reference-count":25,"publisher":"Microbiology Society","issue":"7","license":[{"start":{"date-parts":[[2025,7,3]],"date-time":"2025-07-03T00:00:00Z","timestamp":1751500800000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000923","name":"Australian Research Council","doi-asserted-by":"crossref","award":["IH190100021"],"award-info":[{"award-number":["IH190100021"]}],"id":[{"id":"10.13039\/501100000923","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,7,3]]},"abstract":"<jats:p>\n                  <jats:bold>Introduction.<\/jats:bold>\n                  <jats:italic>Mycoplasma genitalium<\/jats:italic>, a small and slow-growing bacterium, has gained notoriety due to rapidly increasing rates of antibiotic resistance.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Gap Statement.<\/jats:bold>\n                  <jats:italic>M. genitalium<\/jats:italic> is difficult to culture, limiting efforts to understand its biology and the mechanisms of antimicrobial resistance.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Aim.<\/jats:bold> To understand factors that influence success in primary isolation of <jats:italic>M. genitalium<\/jats:italic> from clinical samples.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Methodology.<\/jats:bold> Neat urine or swabs (high vaginal and anal, in universal transport medium) were collected from patients with confirmed or suspected <jats:italic>M. genitalium<\/jats:italic> infections attending the Melbourne Sexual Health Centre. The specimens were stored at \u221280\u2009\u00b0C prior to laboratory analysis. Initial diagnosis was by transcription-mediated amplification (TMA) assay, and samples subsequently testing positive by quantitative PCR (qPCR) were washed twice and inoculated into Vero cell monolayers with a selective antibiotic mixture (cycloheximide and Thayer-Martin Medium I). Cultures were incubated at 37\u2009\u00b0C with 5% CO<jats:sub>2<\/jats:sub> for 8\u2009weeks and observed daily, with qPCR used to monitor growth.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Results.<\/jats:bold> In total, 127 TMA-positive samples were subjected to qPCR, and <jats:italic>M. genitalium<\/jats:italic> genomic DNA (gDNA) was detected in 53.5% (68\/127) of these samples. An isolate was obtained from 26.5% (18\/68) of the gDNA-positive samples following co-culture with Vero cells. The isolation rate varied between sample types, with growth detected in 12.5% (3\/24) of the high vaginal swabs and 37.5% (15\/40) of the urine samples. No isolates were obtained from anal swabs. The proportion with a successful culture was influenced by the initial <jats:italic>M. genitalium<\/jats:italic> load in the sample, which translated into the inoculum size for the Vero cell monolayer. Isolation was unsuccessful with low inoculum (&lt;2,000 genome equivalents, geq), partially successful (13.3%) with a moderate inoculum (2,500\u20139,500 geq) and highly successful (100%) in samples with a high inoculum (&gt;10,000\u2009geq).<\/jats:p>\n               <jats:p>\n                  <jats:bold>Conclusion.<\/jats:bold> The initial bacterial load emerged as a critical determinant of isolation success. This emphasizes the importance of optimizing sample collection and <jats:italic>M. genitalium<\/jats:italic> isolation procedures.<\/jats:p>","DOI":"10.1099\/jmm.0.002040","type":"journal-article","created":{"date-parts":[[2025,7,3]],"date-time":"2025-07-03T19:16:47Z","timestamp":1751570207000},"source":"Crossref","is-referenced-by-count":5,"title":["Factors that contribute to the success of primary isolation of Mycoplasma genitalium from clinical samples"],"prefix":"10.1099","volume":"74","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1321-887X","authenticated-orcid":false,"given":"Jose L.","family":"Huaman","sequence":"first","affiliation":[{"name":"Molecular Microbiology Research Group, Murdoch Children\u2019s Research Institute, Parkville, Victoria, Australia"},{"name":"Department of Obstetrics, Gynaecology and Newborn Health, University of Melbourne, Parkville, Victoria, Australia"},{"name":"Centre for Women\u2019s Infectious Diseases, The Royal Women\u2019s Hospital, Parkville, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6643-5678","authenticated-orcid":false,"given":"Catriona S.","family":"Bradshaw","sequence":"additional","affiliation":[{"name":"Central Clinical School, Monash University, Melbourne, Victoria, Australia"},{"name":"Centre for Epidemiology and Biostatistics, Melbourne School of Population and Global Health, University of Melbourne, Parkville, Victoria, Australia"},{"name":"Melbourne Sexual Health Centre, Alfred Health, Carlton, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7495-6529","authenticated-orcid":false,"given":"Teck-Phui","family":"Chua","sequence":"additional","affiliation":[{"name":"Centre for Women\u2019s Infectious Diseases, The Royal Women\u2019s Hospital, Parkville, Victoria, Australia"},{"name":"Department of Obstetrics, Gynaecology and Newborn Health, University of Melbourne, Parkville, Victoria, Australia"},{"name":"Molecular Microbiology Research Group, Murdoch Children\u2019s Research Institute, Parkville, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5710-2671","authenticated-orcid":false,"given":"Erica L.","family":"Plummer","sequence":"additional","affiliation":[{"name":"Central Clinical School, Monash University, Melbourne, Victoria, Australia"},{"name":"Melbourne Sexual Health Centre, Alfred Health, Carlton, Victoria, Australia"},{"name":"Centre for Women\u2019s Infectious Diseases, The Royal Women\u2019s Hospital, Parkville, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6776-9194","authenticated-orcid":false,"given":"Jennifer A.","family":"Danielewski","sequence":"additional","affiliation":[{"name":"Centre for Women\u2019s Infectious Diseases, The Royal Women\u2019s Hospital, Parkville, Victoria, Australia"},{"name":"Department of Obstetrics, Gynaecology and Newborn Health, University of Melbourne, Parkville, Victoria, Australia"},{"name":"Molecular Microbiology Research Group, Murdoch Children\u2019s Research Institute, Parkville, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3679-9195","authenticated-orcid":false,"given":"Lenka A.","family":"Vodstrcil","sequence":"additional","affiliation":[{"name":"Melbourne Sexual Health Centre, Alfred Health, Carlton, Victoria, Australia"},{"name":"Centre for Epidemiology and Biostatistics, Melbourne School of Population and Global Health, University of Melbourne, Parkville, Victoria, Australia"},{"name":"Central Clinical School, Monash University, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jorgen S.","family":"Jensen","sequence":"additional","affiliation":[{"name":"Research Unit for Reproductive Microbiology, Statens Serum Institut, Copenhagen, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6869-311X","authenticated-orcid":false,"given":"Suzanne M.","family":"Garland","sequence":"additional","affiliation":[{"name":"Centre for Women\u2019s Infectious Diseases, The Royal Women\u2019s Hospital, Parkville, Victoria, Australia"},{"name":"Department of Obstetrics, Gynaecology and Newborn Health, University of Melbourne, Parkville, Victoria, Australia"},{"name":"Molecular Microbiology Research Group, Murdoch Children\u2019s Research Institute, Parkville, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-5012-5659","authenticated-orcid":false,"given":"Natasha","family":"Wild","sequence":"additional","affiliation":[{"name":"Central Clinical School, Monash University, Melbourne, Victoria, Australia"},{"name":"Melbourne Sexual Health Centre, Alfred Health, Carlton, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3330-4433","authenticated-orcid":false,"given":"Gerald L.","family":"Murray","sequence":"additional","affiliation":[{"name":"Department of Obstetrics, Gynaecology and Newborn Health, University of Melbourne, Parkville, Victoria, Australia"},{"name":"Centre for Women\u2019s Infectious Diseases, The Royal Women\u2019s Hospital, Parkville, Victoria, Australia"},{"name":"Molecular Microbiology Research Group, Murdoch Children\u2019s Research Institute, Parkville, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciae295"},{"key":"R2","doi-asserted-by":"publisher","DOI":"10.1016\/j.medj.2024.05.007"},{"key":"R3","doi-asserted-by":"publisher","DOI":"10.1136\/sextrans-2020-054818"},{"key":"R4","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/civ312"},{"key":"R5","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jix145"},{"key":"R6","article-title":"Antibiotic Resistance Threats in the United States, 2019","year":"2019"},{"key":"R7","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(20)30154-7"},{"key":"R8","doi-asserted-by":"publisher","DOI":"10.1016\/j.lanmic.2024.101047"},{"key":"R9","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00006-11"},{"key":"R10","doi-asserted-by":"publisher","DOI":"10.1126\/science.270.5235.397"},{"key":"R11","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.34.2.286-291.1996"},{"key":"R12","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00264-19"},{"key":"R13","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmi.2017.09.006"},{"key":"R14","doi-asserted-by":"publisher","DOI":"10.1016\/j.jiac.2020.11.010"},{"key":"R15","doi-asserted-by":"publisher","DOI":"10.1093\/lambio\/ovad064"},{"key":"R16","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.02056-06"},{"key":"R17","doi-asserted-by":"publisher","DOI":"10.1002\/2052-2975.20"},{"key":"R18","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.49.12.4993-4998.2005"},{"key":"R19","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.42.2.683-692.2004"},{"key":"R20","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkaf174"},{"key":"R21","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkae430"},{"key":"R22","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00819-21"},{"key":"R23","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00344-19"},{"key":"R24","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0162924"},{"key":"R25","doi-asserted-by":"publisher","DOI":"10.4102\/sajid.v36i1.199"}],"container-title":["Journal of Medical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.002040?crawler=true&mimetype=application\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,3]],"date-time":"2025-07-03T19:16:48Z","timestamp":1751570208000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.002040"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,3]]},"references-count":25,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2025,7,3]]}},"URL":"https:\/\/doi.org\/10.1099\/jmm.0.002040","relation":{},"ISSN":["0022-2615","1473-5644"],"issn-type":[{"value":"0022-2615","type":"print"},{"value":"1473-5644","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,3]]}}}