{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T16:27:46Z","timestamp":1768235266781,"version":"3.49.0"},"reference-count":58,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T00:00:00Z","timestamp":1768176000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The overlapping circulation of influenza (Flu), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; SC2), and respiratory syncytial virus (RSV) continues to challenge clinical laboratories, particularly in settings with limited automation and fragmented healthcare coverage. This study expanded the CDC Flu-SC2 assay by incorporating a laboratory-developed test (LDT) for RSV A\/B detection into a fully automated quadruplex RT-qPCR (LDRA) on the Panther Fusion\u00ae Open Access\u2122 system. The design, based on more than 8000 RSV genomic sequences targeting the conserved M gene, achieved optimal amplification efficiencies (97\u2013105%) and full multiplex compatibility. Analytical assessment established limits of detection between 9.6 and 37.8 copies per reaction, absence of cross-reactivity with 30 respiratory pathogens, and inclusivity for 32 viral variants. Commutability and diagnostic performance among the LDRA, CE IVD-marked Allplex\u2122 SARS-CoV-2\/FluA\/FluB\/RSV, and US IVD-marked Panther Fusion\u00ae SARS-CoV-2\/Flu A\/B\/RSV Assays were evaluated using 405 nasopharyngeal UTM-preserved swabs. The LDRA demonstrated excellent concordance (overall agreement \u2265 98%, \u03ba &gt; 0.95), strong diagnostic accuracy, and reliable detection of mixed infections. This quadruplex provides a fully automated, rapid, and accurate solution for the simultaneous detection of influenza A, influenza B, SARS-CoV-2, and RSV viruses, enhancing molecular diagnostic capacity and supporting equitable, timely clinical decision-making in middle-income healthcare systems such as that of the Dominican Republic.<\/jats:p>","DOI":"10.3390\/microorganisms14010167","type":"journal-article","created":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T12:44:44Z","timestamp":1768221884000},"page":"167","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["From Triplex to Quadruplex: Enhancing CDC\u2019s Respiratory qPCR Assay with RSV Detection on Panther Fusion\u00ae Open Access\u2122"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8027-0542","authenticated-orcid":false,"given":"Andy","family":"Caballero M\u00e9ndez","sequence":"first","affiliation":[{"name":"Molecular Biology Department, Referencia Laboratorio Cl\u00ednico, Santo Domingo Oeste, Santo Domingo 11005, Dominican Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-0757-9042","authenticated-orcid":false,"given":"Mayeline N.","family":"Sosa Ortiz","sequence":"additional","affiliation":[{"name":"Molecular Biology Department, Referencia Laboratorio Cl\u00ednico, Santo Domingo Oeste, Santo Domingo 11005, Dominican Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-0877-5396","authenticated-orcid":false,"given":"Roberto A.","family":"Reynoso de la Rosa","sequence":"additional","affiliation":[{"name":"Molecular Biology Department, Referencia Laboratorio Cl\u00ednico, Santo Domingo Oeste, Santo Domingo 11005, Dominican Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-1287-4426","authenticated-orcid":false,"given":"Miguel E.","family":"Abreu Bencosme","sequence":"additional","affiliation":[{"name":"Molecular Biology Department, Referencia Laboratorio Cl\u00ednico, Santo Domingo Oeste, Santo Domingo 11005, Dominican Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-3259-6876","authenticated-orcid":false,"given":"Karla V.","family":"Montero Lebr\u00f3n","sequence":"additional","affiliation":[{"name":"Molecular Biology Department, Referencia Laboratorio Cl\u00ednico, Santo Domingo Oeste, Santo Domingo 11005, Dominican Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.15585\/mmwr.mm6937a6","article-title":"Decreased Influenza Activity During the COVID-19 Pandemic\u2014United States, Australia, Chile, and South Africa, 2020","volume":"69","author":"Olsen","year":"2020","journal-title":"MMWR Morb. 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