{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T17:10:04Z","timestamp":1790097004790,"version":"4.0.1"},"reference-count":43,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institute for Infectious Diseases \u201cLazzaro Spallanzani\u201d"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Arboviral infections represent a growing challenge for global public health due to vector expansion, globalization, and climate change. Dengue (DENV), Zika (ZIKV), Chikungunya (CHIKV), and West Nile virus (WNV) often co-circulate and present overlapping clinical manifestations, making rapid and accurate etiological diagnosis difficult. In this context, multiplex real-time RT-PCR assays can support clinical management and epidemiological surveillance. In this study, analytical and preliminary retrospective comparisons of the NovaplexTM Tropical Fever Virus Assay, a multiplex RT-PCR system designed for the simultaneous detection of DENV, ZIKV, CHIKV, and WNV, were performed. Analytical sensitivity was assessed using serial viral-stock dilutions and Probit regression analysis to estimate the limit of detection (LOD) at a 95% probability. The LOD was 4.41 Log copies\/mL for WNV2, 3.67 for DENV2, 2.57 for ZIKV, and 3.43 for CHIKV in PBS. In spiked plasma or urine, LODs ranged from 2.04 to 4.79 Log copies\/mL. No cross-reactivity with Usutu virus was observed under the conditions tested. In a retrospective comparison of 34 clinical specimens with singleplex assays, agreement was the highest for WNV, substantial for CHIKV, and limited for DENV. These findings support Novaplex as a complementary tool for arboviral diagnosis and surveillance, although further clinical studies are warranted.<\/jats:p>","DOI":"10.3390\/ijms27188398","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T13:32:12Z","timestamp":1790083932000},"page":"8398","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of the NovaplexTM Tropical Fever Virus Assay"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0955-2074","authenticated-orcid":false,"given":"Giuseppe","family":"Sberna","sequence":"first","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0640-8667","authenticated-orcid":false,"given":"Francesca","family":"Colavita","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5022-5388","authenticated-orcid":false,"given":"Maria Beatrice","family":"Valli","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabiano","family":"Brillo","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1952-5117","authenticated-orcid":false,"given":"Fabrizio","family":"Carletti","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0924-993X","authenticated-orcid":false,"given":"Silvia","family":"Meschi","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cosmina","family":"Mija","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7833-9539","authenticated-orcid":false,"given":"Claudia","family":"Minosse","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valentina","family":"Valeriani","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriella","family":"Rozera","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0331-2627","authenticated-orcid":false,"given":"Giulia","family":"Berno","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8975-0089","authenticated-orcid":false,"given":"Flavia","family":"Smoquina","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2975-8611","authenticated-orcid":false,"given":"Lavinia","family":"Fabeni","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7489-5271","authenticated-orcid":false,"given":"Fabrizio","family":"Maggi","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eleonora","family":"Lalle","sequence":"additional","affiliation":[{"name":"Laboratory of Virology and Laboratories of Biosafety, National Institute for Infectious Diseases Lazzaro Spallanzani\u2014IRCCS, 00149 Rome, 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