{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T20:59:23Z","timestamp":1761512363600},"reference-count":28,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2007,4,1]],"date-time":"2007-04-01T00:00:00Z","timestamp":1175385600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2007,4,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Background: Dengue fever is an arthropod-borne infection caused by dengue viruses (DVs; DEN-1 to DEN-4). Early diagnosis is critical to prevent severe disease progression and the spreading of DV because no vaccine or specific treatment is available; therefore, a rapid and specific diagnostic assay capable of detecting and typing all serotypes would be ideal.<\/jats:p>\n               <jats:p>Methods: We amplified RNA samples from all 4 DV serotypes and Japanese encephalitis virus with 4 serotype-specific forward primers and a universal species-specific reverse primer. DEN-1 and DEN-3 forward primers were labeled at their 5\u2032 ends with BODIPY 630\/650 and Cy5.5, respectively. DEN-1 and DEN-3 amplicons were detected by their characteristic emission generated from induced fluorescence resonance energy transfer. The presence of DEN-2 and DEN-4 amplicons was indicated by SYBR Green I (SGI) signals at specific amplicon melting temperatures (Tms).<\/jats:p>\n               <jats:p>Results: Fluorescence signals with specific emission wavelengths were obtained from DEN-1 and DEN-3. SGI melting profiles showed a Tm difference between DEN-2 and DEN-4 of 4.7 \u00b0C, which was sufficient for differentiating these 2 serotypes. The primers did not amplify the Japanese encephalitis virus. The detection limits of DEN-1 to DEN-4 were 1.64 \u00d7 10\u22124, 1.05 \u00d7 10\u22123, 8.15 \u00d7 10\u22124, and 5.80 \u00d7 10\u22123 plaque-forming units per reaction, respectively. The assay had a dynamic range of 103\u2013108 plaque-forming units\/L and could be performed in 2 h.<\/jats:p>\n               <jats:p>Conclusions: A single-tube, 1-step reverse transcription\u2013PCR assay based on Tm and color multiplexing was developed for detecting and typing all 4 DV serotypes.<\/jats:p>","DOI":"10.1373\/clinchem.2006.077446","type":"journal-article","created":{"date-parts":[[2007,2,22]],"date-time":"2007-02-22T20:45:17Z","timestamp":1172177117000},"page":"594-599","source":"Crossref","is-referenced-by-count":10,"title":["One-Step Rapid Reverse Transcription\u2013PCR Assay for Detecting and Typing Dengue Viruses with GC Tail and Induced Fluorescence Resonance Energy Transfer Techniques for Melting Temperature and Color Multiplexing"],"prefix":"10.1093","volume":"53","author":[{"given":"Constance LH","family":"Lo","sequence":"first","affiliation":[{"name":"Department of Health Technology and Informatics, The Hong Kong Polytechnic University, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shea Ping","family":"Yip","sequence":"first","affiliation":[{"name":"Department of Health Technology and Informatics, The Hong Kong Polytechnic University, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter KC","family":"Cheng","sequence":"first","affiliation":[{"name":"Public Health Laboratory Centre, Centre for Health Protection, Department of Health, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tony SS","family":"To","sequence":"first","affiliation":[{"name":"Department of Health Technology and Informatics, The Hong Kong Polytechnic University, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wilina WL","family":"Lim","sequence":"first","affiliation":[{"name":"Public Health Laboratory Centre, Centre for Health Protection, Department of Health, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Polly HM","family":"Leung","sequence":"first","affiliation":[{"name":"Department of Health Technology and Informatics, The Hong Kong Polytechnic University, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2007,4,1]]},"reference":[{"key":"2020022802414124100_R1","doi-asserted-by":"crossref","unstructured":"Moncayo AC, Fernandez Z, Ortiz D, Diallo M, Sall A, Hartman S, et al. 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