{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T07:40:56Z","timestamp":1778658056682,"version":"3.51.4"},"reference-count":32,"publisher":"American Society for Microbiology","issue":"10","license":[{"start":{"date-parts":[[2009,10,1]],"date-time":"2009-10-01T00:00:00Z","timestamp":1254355200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2009,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n            has developed resistance to multiple classes of antimicrobials. There is now growing concern that without the availability of appropriate public health strategies to combat this problem, gonorrhea could become untreatable. For this reason, surveillance for gonococcal antimicrobial resistance must be optimal both in terms of obtaining a representative sample of gonococcal isolates and in terms of having the appropriate tools to identify resistance. To aid with this surveillance, molecular tools are increasingly being used. In the present study, we investigated the use of a simple heat denaturation protocol for isolate DNA preparation combined with SYBR green-based real-time PCR for the identification of mutations associated with\n            <jats:italic>N. gonorrhoeae<\/jats:italic>\n            antimicrobial resistance. A total of 109 clinical gonococcal isolates were tested by high-resolution melting (HRM) curve analysis for chromosomal mutations associated with gonococcal resistance to beta-lactam antibiotics: a\n            <jats:italic>penA<\/jats:italic>\n            345A insertion,\n            <jats:italic>ponA<\/jats:italic>\n            L421P,\n            <jats:italic>mtrR<\/jats:italic>\n            G45D, substitutions at positions 120 and 121 in\n            <jats:italic>porB1b<\/jats:italic>\n            , and an adenine deletion in the\n            <jats:italic>mtrR<\/jats:italic>\n            promoter. An allele-specific PCR assay was also investigated for its ability to detect the adenine deletion in the\n            <jats:italic>mtrR<\/jats:italic>\n            promoter. The results were compared to those obtained by DNA sequencing. Our HRM assays provided the accurate discrimination of heat-treated isolates in which the sequence types differed in GC content, including isolates with the\n            <jats:italic>penA<\/jats:italic>\n            345A insertion and the\n            <jats:italic>ponA<\/jats:italic>\n            L421P and\n            <jats:italic>mtrR<\/jats:italic>\n            G45D mutations. The allele-specific PCR assay accurately identified isolates with the adenine deletion in the\n            <jats:italic>mtrR<\/jats:italic>\n            promoter. Heat-denatured DNA combined with SYBR green-based real-time PCR offers a simple, rapid, and inexpensive means of detecting gonococcal resistance mechanisms. These methods may have broader application in the detection of polymorphisms associated with phenotypes of interest.\n          <\/jats:p>","DOI":"10.1128\/aac.00385-09","type":"journal-article","created":{"date-parts":[[2009,6,16]],"date-time":"2009-06-16T01:12:24Z","timestamp":1245114744000},"page":"4211-4216","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":25,"title":["Simple, Rapid, and Inexpensive Detection of\n            <i>Neisseria gonorrhoeae<\/i>\n            Resistance Mechanisms Using Heat-Denatured Isolates and SYBR Green-Based Real-Time PCR"],"prefix":"10.1128","volume":"53","author":[{"given":"Gayle","family":"Kugelman","sequence":"first","affiliation":[{"name":"Queensland Children's Medical Research Institute, Infectious Diseases Laboratory, Royal Children's Hospital, Herston Road, Herston, QLD 4029, Queensland Australia"},{"name":"Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service District, Herston, Queensland, Australia"},{"name":"Clinical Medical Virology Centre, University of Queensland, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John W.","family":"Tapsall","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for STD and HIV, Microbiology Department, South Eastern Area Laboratory Services, Prince of Wales Hospital, Sydney, New South Wales, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Namraj","family":"Goire","sequence":"additional","affiliation":[{"name":"Queensland Children's Medical Research Institute, Infectious Diseases Laboratory, Royal Children's Hospital, Herston Road, Herston, QLD 4029, Queensland Australia"},{"name":"Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service District, Herston, Queensland, Australia"},{"name":"Clinical Medical Virology Centre, University of Queensland, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Melanie W.","family":"Syrmis","sequence":"additional","affiliation":[{"name":"Queensland Children's Medical Research Institute, Infectious Diseases Laboratory, Royal Children's Hospital, Herston Road, Herston, QLD 4029, Queensland Australia"},{"name":"Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service District, Herston, Queensland, Australia"},{"name":"Clinical Medical Virology Centre, University of Queensland, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Athena","family":"Limnios","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for STD and HIV, Microbiology Department, South Eastern Area Laboratory Services, Prince of Wales Hospital, Sydney, New South Wales, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen B.","family":"Lambert","sequence":"additional","affiliation":[{"name":"Queensland Children's Medical Research Institute, Infectious Diseases Laboratory, Royal Children's Hospital, Herston Road, Herston, QLD 4029, Queensland Australia"},{"name":"Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service District, Herston, Queensland, Australia"},{"name":"Clinical Medical Virology Centre, University of Queensland, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael D.","family":"Nissen","sequence":"additional","affiliation":[{"name":"Queensland Children's Medical Research Institute, Infectious Diseases Laboratory, Royal Children's Hospital, Herston Road, Herston, QLD 4029, Queensland Australia"},{"name":"Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service District, Herston, Queensland, Australia"},{"name":"Clinical Medical Virology Centre, University of Queensland, Queensland, Australia"},{"name":"Microbiology Division, Pathology Queensland, Royal Brisbane Hospital Campus, Brisbane, Queensland, Australia"},{"name":"Department of Paediatric and Child Health, University of Queensland, Brisbane, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Theo P.","family":"Sloots","sequence":"additional","affiliation":[{"name":"Queensland Children's Medical Research Institute, Infectious Diseases Laboratory, Royal Children's Hospital, Herston Road, Herston, QLD 4029, Queensland Australia"},{"name":"Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service District, Herston, Queensland, Australia"},{"name":"Clinical Medical Virology Centre, University of Queensland, Queensland, Australia"},{"name":"Microbiology Division, Pathology Queensland, Royal Brisbane Hospital Campus, Brisbane, Queensland, Australia"},{"name":"Department of Paediatric and Child Health, University of Queensland, Brisbane, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David M.","family":"Whiley","sequence":"additional","affiliation":[{"name":"Queensland Children's Medical Research Institute, Infectious Diseases Laboratory, Royal Children's Hospital, Herston Road, Herston, QLD 4029, Queensland Australia"},{"name":"Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service District, Herston, Queensland, Australia"},{"name":"Clinical Medical Virology Centre, University of Queensland, Queensland, 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