{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T19:18:43Z","timestamp":1791919123624,"version":"4.3.4"},"reference-count":16,"publisher":"Oxford University Press (OUP)","issue":"11","license":[{"start":{"date-parts":[[2022,7,26]],"date-time":"2022-07-26T00:00:00Z","timestamp":1658793600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"name":"Queensland Advancing Clinical Research Fellowship from the Queensland Government"},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000925","name":"NHMRC","doi-asserted-by":"crossref","award":["APP1197743"],"award-info":[{"award-number":["APP1197743"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Advance Queensland Industry Research Fellowship from the Queensland Government"},{"name":"Australian Research Council Research Hub for Antimicrobial Resistance","award":["IH190100021"],"award-info":[{"award-number":["IH190100021"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,10,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>To develop instrument-free point-of-care methods using recombinase polymerase amplification (RPA) technology coupled with a simple lateral flow detection system to detect Neisseria gonorrhoeae and susceptibility to ciprofloxacin.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>For identification of gonococcal infection, an RPA-based method was developed targeting the gonococcal porA pseudogene (NG-porA-RPA). For ciprofloxacin susceptibility, predictive WT sequences at codons 91 and 95 of the gonococcal gyrA DNase gene were targeted. Given the known complexities of SNP detection using RPA (e.g. the ability to accommodate mismatches) we trialled several different assays incorporating various additional non-template mismatches in the oligonucleotide sequences to reduce affinity for the mutant (resistant) gyrA sequences. Assays were evaluated using a bank of N. gonorrhoeae (n\u200a=\u200a10) and non-gonococcal (n\u200a=\u200a5) isolates and a panel of N. gonorrhoeae nucleic acid amplification test (NAAT)-positive clinical sample extracts (n\u200a=\u200a40).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The NG-porA-RPA assay was specific to N. gonorrhoeae and provided a positive percentage agreement (PPA) of 87.5% (35\/40) compared with a commercial N. gonorrhoeae NAAT when applied to the 40 clinical sample extracts. For gyrA, the non-template bases successfully reduced banding intensity for double-mutant strains (mutations at both 91 and 95), but not for rarer single-mutant (91 only) strains. The most promising gyrA assay, NG-gyrA-RPA08, correctly detected 83% (25\/30) of infections from NAAT-positive clinical samples confirmed to have WT gyrA sequences based on Sanger sequencing.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>These proof-of-concept data show that RPA technology has considerable promise for detecting N. gonorrhoeae and associated antibiotic susceptibility and would offer a diagnostic-based stewardship strategy identified as urgently needed by the WHO.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/jac\/dkac242","type":"journal-article","created":{"date-parts":[[2022,7,26]],"date-time":"2022-07-26T07:48:42Z","timestamp":1658821722000},"page":"2933-2936","source":"Crossref","is-referenced-by-count":10,"title":["Proof-of-concept, rapid, instrument-free molecular detection of\n                    <i>Neisseria gonorrhoeae<\/i>\n                    and ciprofloxacin susceptibility"],"prefix":"10.1093","volume":"77","author":[{"given":"Abdulrahman K S","family":"Ayfan","sequence":"first","affiliation":[{"name":"Faculty of Medicine, The University of Queensland Centre for Clinical Research (UQCCR) , Brisbane , Australia"},{"name":"Faculty of Science, Biochemistry Department, King Abdul-Aziz University (KAU) , Jeddah , Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanne","family":"Macdonald","sequence":"additional","affiliation":[{"name":"School of Science and Engineering, Genecology Research Centre, University of Sunshine Coast (USC) , Sunshine Coast , Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8974-6789","authenticated-orcid":false,"given":"Adam D","family":"Irwin","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, The University of Queensland Centre for Clinical Research (UQCCR) , Brisbane , Australia"},{"name":"Infection Management and Prevention Services, Queensland Children\u2019s Hospital , Brisbane , Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hosam M","family":"Zowawi","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, The University of Queensland Centre for Clinical Research (UQCCR) , Brisbane , Australia"},{"name":"College of Medicine, King Saud bin Abdulaziz University for Health Sciences , Riyadh , Saudi Arabia"},{"name":"King Abdullah International Medical Research Centre , Riyadh , Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2264-4785","authenticated-orcid":false,"given":"Brian M","family":"Forde","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, The University of Queensland Centre for Clinical Research (UQCCR) , Brisbane , Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2079-4437","authenticated-orcid":false,"given":"David L","family":"Paterson","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, The University of Queensland Centre for Clinical Research (UQCCR) , Brisbane , Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica M","family":"Lahra","sequence":"additional","affiliation":[{"name":"World Health Organization Collaborating Centre for STDs, Prince of Wales Hospital , Sydney , Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David M","family":"Whiley","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, The University of Queensland Centre for Clinical Research (UQCCR) , Brisbane , Australia"},{"name":"Infection Management and Prevention Services, Queensland Children\u2019s Hospital , Brisbane , Australia"},{"name":"Pathology Queensland, Department of Microbiology , Brisbane , Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,7,26]]},"reference":[{"key":"2022102819180750000_dkac242-B1","author":"WHO","year":"2017"},{"key":"2022102819180750000_dkac242-B2","author":"CDC","year":"2019"},{"key":"2022102819180750000_dkac242-B3","doi-asserted-by":"crossref","first-page":"1900147","DOI":"10.2807\/1560-7917.ES.2019.24.10.1900147","article-title":"Detection in the United Kingdom of the Neisseria gonorrhoeae FC428 clone, with ceftriaxone resistance and intermediate resistance to azithromycin, October to December 2018","volume":"24","author":"Eyre","year":"2019","journal-title":"Euro Surveill"},{"key":"2022102819180750000_dkac242-B4","doi-asserted-by":"crossref","first-page":"1900118","DOI":"10.2807\/1560-7917.ES.2019.24.8.1900118","article-title":"Genetic relatedness of ceftriaxone-resistant and high-level azithromycin resistant Neisseria gonorrhoeae cases, United Kingdom 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