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Research Lecturer Grant, Faculty of Medicine Siriraj Hospital, Mahidol University","award":["5680049"],"award-info":[{"award-number":["5680049"]}]},{"name":"a Research Lecturer Grant, Faculty of Medicine Siriraj Hospital, Mahidol University","award":["PHD\/0101\/2554"],"award-info":[{"award-number":["PHD\/0101\/2554"]}]},{"name":"the Research Excellence Development (RED) program, Faculty of Medicine Siriraj Hospital, Mahidol University","award":["R015834004"],"award-info":[{"award-number":["R015834004"]}]},{"name":"the Research Excellence Development (RED) program, Faculty of Medicine Siriraj Hospital, Mahidol University","award":["5680049"],"award-info":[{"award-number":["5680049"]}]},{"name":"the Research Excellence Development (RED) program, Faculty of Medicine Siriraj Hospital, Mahidol University","award":["PHD\/0101\/2554"],"award-info":[{"award-number":["PHD\/0101\/2554"]}]},{"name":"Research Chair Grant, NSTDA, Thailand","award":["R015834004"],"award-info":[{"award-number":["R015834004"]}]},{"name":"Research Chair Grant, NSTDA, Thailand","award":["5680049"],"award-info":[{"award-number":["5680049"]}]},{"name":"Research Chair Grant, NSTDA, Thailand","award":["PHD\/0101\/2554"],"award-info":[{"award-number":["PHD\/0101\/2554"]}]},{"name":"Ph.D. Scholar in the Royal Golden Jubilee Ph.D. Program","award":["R015834004"],"award-info":[{"award-number":["R015834004"]}]},{"name":"Ph.D. Scholar in the Royal Golden Jubilee Ph.D. Program","award":["5680049"],"award-info":[{"award-number":["5680049"]}]},{"name":"Ph.D. Scholar in the Royal Golden Jubilee Ph.D. Program","award":["PHD\/0101\/2554"],"award-info":[{"award-number":["PHD\/0101\/2554"]}]},{"name":"Graduate Scholarship from Faculty of Medicine Siriraj Hospital, Mahidol University","award":["R015834004"],"award-info":[{"award-number":["R015834004"]}]},{"name":"Graduate Scholarship from Faculty of Medicine Siriraj Hospital, Mahidol University","award":["5680049"],"award-info":[{"award-number":["5680049"]}]},{"name":"Graduate Scholarship from Faculty of Medicine Siriraj Hospital, Mahidol University","award":["PHD\/0101\/2554"],"award-info":[{"award-number":["PHD\/0101\/2554"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Dengue virus (DENV) infection is a significant global health problem. There are no specific therapeutics or widely available vaccines. Early diagnosis is critical for patient management. Viral RNA detection by multiplex RT-PCR using multiple pairs of primers\/probes allowing the simultaneous detection of all four DENV serotypes is commonly used. However, increasing the number of primers in the RT-PCR reaction reduces the sensitivity of detection due to the increased possibility of primer dimer formation. Here, a one tube, singleplex real-time RT-PCR specific to DENV 3\u2032-UTR was developed for the detection and quantification of pan-DENV with no cross reactivity to other flaviviruses. The sensitivity of DENV detection was as high as 96.9% in clinical specimens collected at the first day of hospitalization. Our assay provided equivalent PCR efficiency and RNA quantification among each DENV serotype. The assay\u2019s performance was comparable with previously established real-time RT-PCR targeting coding sequences. Using both assays on the same specimens, our results indicate the presence of defective virus particles in the circulation of patients infected with all serotypes. Dual regions targeting RT-PCR enhanced the sensitivity of viral genome detection especially during the late acute phase when viremia rapidly decline and an incomplete viral genome was clinically evident.<\/jats:p>","DOI":"10.3390\/v14061271","type":"journal-article","created":{"date-parts":[[2022,6,13]],"date-time":"2022-06-13T06:31:59Z","timestamp":1655101919000},"page":"1271","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Development of a Singleplex Real-Time Reverse Transcriptase PCR Assay for Pan-Dengue Virus Detection and Quantification"],"prefix":"10.3390","volume":"14","author":[{"given":"Adisak","family":"Songjaeng","sequence":"first","affiliation":[{"name":"Division of Dengue Hemorrhagic Fever Research, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Somchai","family":"Thiemmeca","sequence":"additional","affiliation":[{"name":"Division of Dengue Hemorrhagic Fever Research, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dumrong","family":"Mairiang","sequence":"additional","affiliation":[{"name":"Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Bangkok 12120, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nuntaya","family":"Punyadee","sequence":"additional","affiliation":[{"name":"Division of Dengue Hemorrhagic Fever Research, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5937-7254","authenticated-orcid":false,"given":"Kessiri","family":"Kongmanas","sequence":"additional","affiliation":[{"name":"Division of Dengue Hemorrhagic Fever Research, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prachya","family":"Hansuealueang","sequence":"additional","affiliation":[{"name":"Graduate Program in Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nattaya","family":"Tangthawornchaikul","sequence":"additional","affiliation":[{"name":"Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Bangkok 12120, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thaneeya","family":"Duangchinda","sequence":"additional","affiliation":[{"name":"Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Bangkok 12120, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juthathip","family":"Mongkolsapaya","sequence":"additional","affiliation":[{"name":"Wellcome Centre for Human Genetics, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7BN, UK"},{"name":"Chinese Academy of Medical Science (CAMS) Oxford Institute (COI), University of Oxford, Oxford OX3 7FZ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kanokwan","family":"Sriruksa","sequence":"additional","affiliation":[{"name":"Pediatric Department, Khon Kaen Hospital, Ministry of Public Health, Khon Kaen 40000, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wannee","family":"Limpitikul","sequence":"additional","affiliation":[{"name":"Pediatric Department, Songkhla Hospital, Ministry of Public Health, Songkhla 90100, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prida","family":"Malasit","sequence":"additional","affiliation":[{"name":"Division of Dengue Hemorrhagic Fever Research, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Bangkok 12120, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0053-3045","authenticated-orcid":false,"given":"Panisadee","family":"Avirutnan","sequence":"additional","affiliation":[{"name":"Division of Dengue Hemorrhagic Fever Research, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok 10700, Thailand"},{"name":"Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Bangkok 12120, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1038\/nature12060","article-title":"The global distribution and burden of dengue","volume":"496","author":"Bhatt","year":"2013","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1508","DOI":"10.1038\/s41564-019-0476-8","article-title":"The current and future global distribution and population at risk of dengue","volume":"4","author":"Messina","year":"2019","journal-title":"Nat. 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