{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T07:04:31Z","timestamp":1790665471180,"version":"4.1.0"},"reference-count":16,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2021,1,15]],"date-time":"2021-01-15T00:00:00Z","timestamp":1610668800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Pathology Queensland Study Education and Research Committee","award":["SERC5920"],"award-info":[{"award-number":["SERC5920"]}]},{"name":"Queensland Advancing Clinical Research Fellowship"},{"DOI":"10.13039\/501100003550","name":"Queensland Government","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003550","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,3,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Mycoplasma genitalium was recently added to the CDC\u2019s antimicrobial resistance threats \u2018watch list\u2019, as it has rapidly become resistant to mainstay treatments. In Australia, treatment failure with fluoroquinolones remain commonplace, even when Sanger sequencing fails to identify evidence of resistance mutations.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Suspecting that Sanger sequencing may miss low-load mixed infections, we applied three additional PCR-based approaches (allele-specific primer-based PCR, probe-based PCR and amplicon deep sequencing) to detect mutations associated with fluoroquinolone susceptibility\/resistance. We focused on resistance mutations at amino acid positions 83 and 87 of parC, as these were previously shown to be common in Australia.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Our results showed evidence of mixtures of fluoroquinolone-susceptible and -resistant strains in up to 27\/423 samples (6.4%). These included 1 sample that was indicated to be mixed by Sanger sequencing and all three additional PCR methods, 6 samples detected by two or more of the additional PCRs but not by Sanger sequencing and finally 20 samples that were detected by only one of the additional PCR methods. A key question was whether Sanger sequencing failed to detect fluoroquinolone resistance in any samples; overall, we observed that Sanger sequencing failed to detect fluoroquinolone resistance in up to 3.8% (16\/423) of samples.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>The presence of mixed susceptibility infections may have important implications for clinical patient management and stresses the need for appropriate detection of resistance and selection of antimicrobials to ensure appropriate treatment of M. genitalium infections.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/jac\/dkaa542","type":"journal-article","created":{"date-parts":[[2020,12,15]],"date-time":"2020-12-15T17:44:19Z","timestamp":1608054259000},"page":"887-892","source":"Crossref","is-referenced-by-count":12,"title":["<i>Mycoplasma genitalium<\/i> infections can comprise a mixture of both fluoroquinolone-susceptible and fluoroquinolone-resistant strains"],"prefix":"10.1093","volume":"76","author":[{"given":"Emma L","family":"Sweeney","sequence":"first","affiliation":[{"name":"The University of Queensland Centre for Clinical Research (UQ-CCR), Faculty of Medicine, The University of Queensland, Brisbane, Queensland, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kym","family":"Lowry","sequence":"additional","affiliation":[{"name":"The University of Queensland Centre for Clinical Research (UQ-CCR), Faculty of Medicine, The University of Queensland, Brisbane, Queensland, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheryl","family":"Bletchly","sequence":"additional","affiliation":[{"name":"Pathology Queensland Central Laboratory, Queensland, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Graeme R","family":"Nimmo","sequence":"additional","affiliation":[{"name":"Griffith University, School of Medicine, Queensland, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David M","family":"Whiley","sequence":"additional","affiliation":[{"name":"The University of Queensland Centre for Clinical Research (UQ-CCR), Faculty of Medicine, The University of Queensland, Brisbane, Queensland, Australia"},{"name":"Pathology Queensland Central Laboratory, Queensland, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,1,15]]},"reference":[{"key":"2021031209291429700_dkaa542-B1","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1128\/CMR.00006-11","article-title":"Mycoplasma genitalium: from chrysalis to multicolored butterfly","volume":"24","author":"Taylor-Robinson","year":"2011","journal-title":"Clin Microbiol Rev"},{"key":"2021031209291429700_dkaa542-B2","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/j.diagmicrobio.2016.07.005","article-title":"Emergence of Mycoplasma genitalium strains showing mutations associated with macrolide and fluoroquinolone resistance in the region Dresden, Germany","volume":"86","author":"Dumke","year":"2016","journal-title":"Diagn Microbiol Infect Dis"},{"key":"2021031209291429700_dkaa542-B3","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.3201\/eid2209.160446","article-title":"Fluoroquinolone-resistant Mycoplasma genitalium, southwestern France","volume":"22","author":"Le Roy","year":"2016","journal-title":"Emerg Infect Dis"},{"key":"2021031209291429700_dkaa542-B4","doi-asserted-by":"crossref","first-page":"e01555-18","DOI":"10.1128\/JCM.01555-18","article-title":"Levels of Mycoplasma genitalium antimicrobial resistance differ by both region and gender in the state of Queensland, Australia: implications for treatment guidelines","volume":"57","author":"Sweeney","year":"2019","journal-title":"J Clin Microbiol"},{"key":"2021031209291429700_dkaa542-B5","doi-asserted-by":"crossref","first-page":"1302","DOI":"10.1016\/S1473-3099(20)30154-7","article-title":"Prevalence of mutations associated with resistance to macrolides and fluoroquinolones in Mycoplasma genitalium: a systematic review and meta-analysis","volume":"20","author":"Machalek","year":"2020","journal-title":"Lancet Infect Dis"},{"key":"2021031209291429700_dkaa542-B6","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1016\/j.jiac.2018.08.009","article-title":"Surveillance of the prevalence of macrolide and\/or fluoroquinolone resistance-associated mutations in Mycoplasma genitalium in Japan","volume":"24","author":"Deguchi","year":"2018","journal-title":"J Infect Chemother"},{"key":"2021031209291429700_dkaa542-B7","doi-asserted-by":"crossref","first-page":"e00886-19","DOI":"10.1128\/JCM.00886-19","article-title":"Multicenter clinical evaluation of a novel multiplex real-time PCR (qPCR) assay for detection of fluoroquinolone resistance in Mycoplasma genitalium","volume":"57","author":"Fern\u00e1ndez-Huerta","year":"2019","journal-title":"J Clin Microbiol"},{"key":"2021031209291429700_dkaa542-B8","doi-asserted-by":"crossref","first-page":"2245","DOI":"10.1128\/JCM.00495-13","article-title":"Fluoroquinolone and macrolide resistance-associated mutations in Mycoplasma genitalium","volume":"51","author":"Tagg","year":"2013","journal-title":"J Clin Microbiol"},{"key":"2021031209291429700_dkaa542-B9","doi-asserted-by":"crossref","first-page":"4211","DOI":"10.1128\/AAC.00385-09","article-title":"Simple, rapid, and inexpensive detection of Neisseria gonorrhoeae resistance mechanisms using heat-denatured isolates and SYBR green-based real-time PCR","volume":"53","author":"Kugelman","year":"2009","journal-title":"Antimicrob Agents Chemother"},{"key":"2021031209291429700_dkaa542-B10","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1186\/1471-2180-8-130","article-title":"Short tandem repeat sequences in the Mycoplasma genitalium genome and their use in a multilocus genotyping system","volume":"8","author":"Ma","year":"2008","journal-title":"BMC Microbiol"},{"key":"2021031209291429700_dkaa542-B11","doi-asserted-by":"crossref","first-page":"e00628-17","DOI":"10.1128\/AAC.00628-17","article-title":"Digital PCR for detection and quantification of fluoroquinolone resistance in Legionella pneumophila","volume":"61","author":"Hennebique","year":"2017","journal-title":"Antimicrob Agents Chemother"},{"key":"2021031209291429700_dkaa542-B12","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1093\/jac\/dkw406","article-title":"Mixed gonococcal infections in a high-risk population, Sydney, Australia 2015: implications for antimicrobial resistance surveillance?","volume":"72","author":"Goire","year":"2017","journal-title":"J Antimicrob Chemother"},{"key":"2021031209291429700_dkaa542-B13","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.meegid.2015.10.028","article-title":"Multiple and mixed Helicobacter pylori infections: comparison of two epidemiological situations in Tunisia and France","volume":"37","author":"Ben Mansour","year":"2016","journal-title":"Infect Genet Evol"},{"key":"2021031209291429700_dkaa542-B14","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.anaerobe.2018.05.017","article-title":"Presence of multiple Clostridium difficile strains at primary infection is associated with development of recurrent disease","volume":"53","author":"Seekatz","year":"2018","journal-title":"Anaerobe"},{"key":"2021031209291429700_dkaa542-B15","doi-asserted-by":"crossref","first-page":"S412","DOI":"10.1093\/infdis\/jix132","article-title":"New horizons in Mycoplasma genitalium treatment","volume":"216","author":"Bradshaw","year":"2017","journal-title":"J Infect Dis"},{"key":"2021031209291429700_dkaa542-B16","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1097\/OLQ.0000000000001132","article-title":"Quinolone resistance-associated mutations in Mycoplasma genitalium: not ready for prime time","volume":"47","author":"Manhart","year":"2020","journal-title":"Sex Transm Dis"}],"container-title":["Journal of Antimicrobial Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/jac\/article-pdf\/76\/4\/887\/36580549\/dkaa542.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/jac\/article-pdf\/76\/4\/887\/36580549\/dkaa542.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T09:38:24Z","timestamp":1615541904000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jac\/article\/76\/4\/887\/6100778"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,15]]},"references-count":16,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,1,15]]},"published-print":{"date-parts":[[2021,3,12]]}},"URL":"https:\/\/doi.org\/10.1093\/jac\/dkaa542","relation":{},"ISSN":["0305-7453","1460-2091"],"issn-type":[{"value":"0305-7453","type":"print"},{"value":"1460-2091","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2021,4,1]]},"published":{"date-parts":[[2021,1,15]]}}}