{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T13:53:56Z","timestamp":1777298036169,"version":"3.51.4"},"reference-count":16,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T00:00:00Z","timestamp":1764892800000},"content-version":"vor","delay-in-days":338,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,1,1]],"date-time":"2025-01-01T00:00:00Z","timestamp":1735689600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100004410","name":"T\u00fcrkiye Bilimsel ve Teknolojik Ara\u015ftirma Kurumu","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004410","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Canadian Journal of Infectious Diseases and Medical Microbiology"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Recent advances in detecting respiratory pathogens have allowed for the simultaneous identification of multiple agents, enabling quick and accurate diagnosis to start timely treatment. This study aimed to design a novel two\u2010dimensional (2D) multiplex reverse transcription quantitative PCR (RT\u2010qPCR) assay. This assay allows for the concurrent detection of SARS\u2010CoV\u20102, RSV, and Influenza A\u2010B using molecular beacon technology in a single\u2010well format.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We used 550 nasopharyngeal swab samples from the Kocaeli University, Research and Educational Hospital, PCR laboratory, along with synthetic plasmids for SARS\u2010CoV\u20102, RSV, Influenza A\u2010B, and internal control (RNase P). DNA products generated after amplification interacted with intermediate probes containing specific enzymatic cleavage sites and fluorescent markers, producing characteristic melting temperature (Tm) values for melting curve analysis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Distinct Tm values were identified for SARS\u2010CoV\u20102 (72\u00b0C), RSV (66\u00b0C), Influenza A (56\u00b0C), Influenza B (68\u00b0C), and internal control (80.5\u00b0C). The accuracy was confirmed by testing laboratory\u2010confirmed samples and synthetic plasmids, with no cross\u2010reactivity or false positives observed.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This melting curve\u2013based assay can differentiate among various pathogens within a single well and fluorescence channel by utilizing the unique Tm of each target. Consequently, this novel assay may serve as a cost\u2010effective, high\u2010throughput PCR testing method compared to traditional diagnostics.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1155\/cjid\/6687705","type":"journal-article","created":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T12:37:40Z","timestamp":1764938260000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Design of a 2D Melting Curve\u2013Based Multiplex PCR Assay for Detection of SARS\u2010CoV\u20102\/RSV\/Influenza A\u2010B"],"prefix":"10.1155","volume":"2025","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4374-7193","authenticated-orcid":false,"given":"Murat","family":"Sayan","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1942-1203","authenticated-orcid":false,"given":"Ayse","family":"Arikan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8904-7064","authenticated-orcid":false,"given":"Tahaberk","family":"Ateso\u011flu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,12,5]]},"reference":[{"key":"e_1_2_11_1_2","unstructured":"The World Health Organisation (WHO) Global Health Estimates: Life Expectancy and Leading Causes of Death and Disability https:\/\/www.who.int\/data\/gho\/data\/themes\/mortality-and-global-health-estimates."},{"key":"e_1_2_11_2_2","unstructured":"European Center for Disease Prevention and Control (ECDC) Weekly respiratory virus updates week 4 January 2025 https:\/\/www.ecdc.europa.eu\/en\/publications-data\/weekly-respiratory-virus-update-week-4-january-2025."},{"key":"e_1_2_11_3_2","unstructured":"New York City Department of Health and Mental Hygiene Influenza and RSV Surveillance Report 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Journal of Infectious Diseases and Medical 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