{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T17:58:31Z","timestamp":1791568711672,"version":"4.3.4"},"reference-count":42,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2023,3,6]],"date-time":"2023-03-06T00:00:00Z","timestamp":1678060800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Vietnam National University, Hanoi and the Ministry of Science and Technology, Vietnam","award":["KLEPT.20.04"],"award-info":[{"award-number":["KLEPT.20.04"]}]},{"name":"Vingroup JSC and Vingroup Innovation Foundation (VINIF), Institute of Big Data","award":["VINIF.2021.ThS.72"],"award-info":[{"award-number":["VINIF.2021.ThS.72"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Sexually transmitted diseases are major causes of infertility, ectopic pregnancy, and premature birth. Here, we developed a new multiplex real-time polymerase chain reaction (PCR) assay for the simultaneous detection of nine major sexually transmitted infections (STIs) found in Vietnamese women, including <jats:italic>Chlamydia trachomatis<\/jats:italic>, <jats:italic>Neisseria gonorrhoeae<\/jats:italic>, <jats:italic>Gardnerella vaginalis<\/jats:italic>, <jats:italic>Trichomonas vaginalis<\/jats:italic>, <jats:italic>Candida albicans<\/jats:italic>, <jats:italic>Mycoplasma hominis<\/jats:italic>, <jats:italic>Mycoplasma genitalium<\/jats:italic>, and human alphaherpesviruses 1 and 2. A panel containing three tubes \u00d7 three pathogens\/tube was predesigned based on double-quenched TaqMan probes to increase detection sensitivity. There was no cross-reactivity among the nine STIs and other non-targeted microorganisms. Depending on each pathogen, the agreement with commercial kits, sensitivity, specificity, repeatability and reproducibility coefficient of variation (CV), and limit of detection of the developed real-time PCR assay were 99.0%\u2013100%, 92.9%\u2013100%, 100%, &lt;3%, and 8\u201358 copies\/reaction, respectively. One assay cost only 2.34 USD. Application of the assay for the detection of the nine STIs in 535 vaginal swab samples collected from women in Vietnam yielded 532 positive cases (99.44%). Among the positive samples, 37.76% had one pathogen, with <jats:italic>G<\/jats:italic>. <jats:italic>vaginalis<\/jats:italic> (33.83%) as the most prevalent; 46.36% had two pathogens, with <jats:italic>G<\/jats:italic>. <jats:italic>vaginalis<\/jats:italic> + <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> as the most prevalent combination (38.13%); and 11.78%, 2.99%, and 0.56% had three, four, and five pathogens, respectively. In conclusion, the developed assay represents a sensitive and cost-effective molecular diagnostic tool for the detection of major STIs in Vietnam and is a model for the development of panel detections of common STIs in other countries.<\/jats:p>","DOI":"10.1371\/journal.pone.0282439","type":"journal-article","created":{"date-parts":[[2023,3,6]],"date-time":"2023-03-06T18:25:25Z","timestamp":1678127125000},"page":"e0282439","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":22,"title":["Simultaneous real-time PCR detection of nine prevalent sexually transmitted infections using a predesigned double-quenched TaqMan probe panel"],"prefix":"10.1371","volume":"18","author":[{"given":"Ha T. V.","family":"Bui","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huyen T.","family":"Bui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Son V.","family":"Chu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huyen T.","family":"Nguyen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1215-6910","authenticated-orcid":true,"given":"Anh T. V.","family":"Nguyen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Phuong T.","family":"Truong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thang T. H.","family":"Dang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anh T. 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Available from: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/sexually-transmitted-infections-(stis)."},{"issue":"10","key":"pone.0282439.ref002","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1097\/OLQ.0b013e318265b180","article-title":"Prevalence of HIV\/STIs and associated factors among men who have sex with men in An Giang, Vietnam","volume":"39","author":"QD Pham","year":"2012","journal-title":"Sex Transm Dis"},{"issue":"2","key":"pone.0282439.ref003","first-page":"133","article-title":"Acceptability and feasibility of sexually transmissible infection screening among pregnant women in Hanoi, Vietnam","volume":"16","author":"M Nguyen","year":"2019","journal-title":"Sex Health"},{"issue":"2","key":"pone.0282439.ref004","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1177\/09564624211060185","article-title":"HIV and STI prevalence and testing history among men who have sex with men in Hanoi, Vietnam","volume":"33","author":"JS 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isolation in cell culture","volume":"188","author":"A Wald","year":"2003","journal-title":"J Infect Dis"},{"issue":"3","key":"pone.0282439.ref008","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1080\/10647440500068248","article-title":"Real-time PCR improves detection of Trichomonas vaginalis infection compared with culture using self-collected vaginal swabs","volume":"13","author":"AM Caliendo","year":"2005","journal-title":"Infect Dis Obstet Gynecol"},{"issue":"5","key":"pone.0282439.ref009","doi-asserted-by":"crossref","first-page":"2058","DOI":"10.1128\/JCM.43.5.2058-2064.2005","article-title":"Detection and typing of Herpes simplex virus (HSV) in mucocutaneous samples by TaqMan PCR targeting a gB segment homologous for HSV types 1 and 2","volume":"43","author":"L Namvar","year":"2005","journal-title":"J Clin Microbiol"},{"issue":"1","key":"pone.0282439.ref010","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1128\/JCM.44.1.206-213.2006","article-title":"Development and validation of a rotor-gene real-time PCR assay for detection, identification, and quantification of Chlamydia trachomatis in a single reaction","volume":"44","author":"H Jalal","year":"2006","journal-title":"J Clin Microbiol"},{"issue":"1","key":"pone.0282439.ref011","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.diagmicrobio.2007.12.007","article-title":"A duplex Neisseria gonorrhoeae real-time polymerase chain reaction assay targeting the gonococcal porA pseudogene and multicopy opa genes","volume":"61","author":"N Goire","year":"2008","journal-title":"Diagn Microbiol Infect Dis"},{"issue":"7","key":"pone.0282439.ref012","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1099\/jmm.0.009977-0","article-title":"Real-time PCR detection of the mg219 gene of unknown function of Mycoplasma 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Trembizki","year":"2015","journal-title":"J Antimicrob Chemother"},{"issue":"9","key":"pone.0282439.ref016","doi-asserted-by":"crossref","first-page":"3236","DOI":"10.1128\/JCM.01019-10","article-title":"Performance of the Abbott Real-time CT\/NG for detection of Chlamydia trachomatis and Neisseria gonorrhoeae","volume":"48","author":"CA Gaydos","year":"2010","journal-title":"J Clin Microbiol"},{"issue":"10","key":"pone.0282439.ref017","doi-asserted-by":"crossref","first-page":"3544","DOI":"10.1128\/JCM.38.10.3544-3549.2000","article-title":"Multicenter evaluation of Amplicor and automated Cobas Amplicor CT\/NG tests for Neisseria gonorrhoeae","volume":"38","author":"DH Martin","year":"2000","journal-title":"J Clin Microbiol"},{"issue":"3","key":"pone.0282439.ref018","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1128\/JCM.38.3.1105-1112.2000","article-title":"Multicenter evaluation of the Aplicor and automated Cobas Amplicor CT\/NG tests for detection of Chlamydia 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Neisseria gonorrhoeae","volume":"71","author":"V Doseeva","year":"2011","journal-title":"Diagn Microbiol Infect Dis"},{"issue":"2","key":"pone.0282439.ref027","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1016\/j.yexmp.2015.01.011","article-title":"A multiplex PCR assay for the simultaneous detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis","volume":"98","author":"AN Abou Tayoun","year":"2015","journal-title":"Exp Mol Pathol"},{"issue":"1","key":"pone.0282439.ref028","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/s12879-018-3629-0","article-title":"Comparison of an in-house real-time duplex PCR assay with commercial Hologic\u00ae Aptima assays for the detection of Neisseria gonorrhoeae and Chlamydia trachomatis in urine and extra-genital specimens","volume":"19","author":"JME Venter","year":"2019","journal-title":"BMC Infect Dis"},{"key":"pone.0282439.ref029","doi-asserted-by":"crossref","first-page":"382","DOI":"10.3389\/fcimb.2019.00382","article-title":"Design and evaluation of a novel multiplex real-time PCR melting curve assay for the simultaneous detection of nine sexually transmitted disease pathogens in genitourinary secretions","volume":"9","author":"XM Hu","year":"2019","journal-title":"Front Cell Infect Microbiol"},{"issue":"2","key":"pone.0282439.ref030","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/j.mimet.2011.12.017","article-title":"Development of a rotor-gene real-time PCR assay for the detection and quantification of Mycoplasma genitalium","volume":"88","author":"EE Muller","year":"2012","journal-title":"J Microbiol Methods"},{"issue":"1","key":"pone.0282439.ref031","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1186\/s12879-020-05066-8","article-title":"Molecular epidemiology and socio-demographic risk factors of sexually transmitted infections among women in Lebanon","volume":"20","author":"J Hanna","year":"2020","journal-title":"BMC Infect Dis"},{"key":"pone.0282439.ref032","first-page":"2020","article-title":"Risk factors associated with prevalence of Candida albicans, Gardnerella vaginalis, and Trichomonas vaginalis among women at the district hospital of Dschang, West Region","author":"VK Payne","year":"2020","journal-title":"Cameroon. Int J Microbiol"},{"issue":"1","key":"pone.0282439.ref033","doi-asserted-by":"crossref","first-page":"73","DOI":"10.30574\/gscbps.2022.18.1.0026","article-title":"Prevalence of Candida albicans species among females with symptoms","volume":"18","author":"U Fajoyomi Bridget","year":"2022","journal-title":"GSC Biol Pharm Sci"},{"issue":"4","key":"pone.0282439.ref034","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1016\/j.jpag.2018.01.003","article-title":"Prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis infection in Chilean adolescents and young adults","volume":"31","author":"A Huneeus","year":"2018","journal-title":"J Pediatr Adolesc Gynecol"},{"key":"pone.0282439.ref035","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1186\/s12879-015-0792-4","article-title":"Prevalence of Mycoplasma genitalium and Mycoplasma hominis in urogenital tract of Brazilian women","volume":"15","author":"GB Campos","year":"2015","journal-title":"BMC Infect Dis"},{"key":"pone.0282439.ref036","article-title":"Infection prevalence and antibiotic resistance levels in Ureaplasma urealyticum and Mycoplasma hominis in gynecological outpatients of a Tertiary hospital in China from 2015 to 2018","author":"W Zhang","year":"2021","journal-title":"Can J Infect Dis Med Microbiol"},{"issue":"5","key":"pone.0282439.ref037","doi-asserted-by":"crossref","first-page":"1271","DOI":"10.3349\/ymj.2016.57.5.1271","article-title":"Prevalence and antibiotic susceptibility of Mycoplasma hominis and Ureaplasma urealyticum in pregnant women","volume":"57","author":"MY Lee","year":"2016","journal-title":"Yonsei Med J"},{"issue":"3","key":"pone.0282439.ref038","doi-asserted-by":"crossref","DOI":"10.3390\/medicina57030211","article-title":"The prevalence of Ureaplasma urealyticum and Mycoplasma hominis infections in infertile patients in the Northeast Region of Romania","volume":"57","author":"B Doroftei","year":"2021","journal-title":"Medicina"},{"issue":"3","key":"pone.0282439.ref039","doi-asserted-by":"crossref","first-page":"180","DOI":"10.4274\/jus.galenos.2019.2405","article-title":"Prevalence of polymicrobial infection in urethritis","volume":"6","author":"M. 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