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Serological diagnosis of these viruses is challenging, making the development of new diagnostic strategies imperative. In this study, we investigated the effectiveness of gold nanorods (GNRs) functionalized with specific anti-dengue and anti-orthoflavivirus antibodies in detecting viral particles. GNRs were created with a length-to-width ratio of up to 5.5, a size of 71.4 \u00b1 6.5 nm, and a light absorption peak at 927 nm, and they were treated with 4 mM polyethyleneimine. These GNRs were attached to a small amount of monoclonal antibodies that target flaviviruses, and the viral particles were detected by measuring the localized surface plasmon resonance using an UV-Vis\/NIR spectrometer. The tests found Orthoflavivirus dengue and Orthoflavivirus zikaense in diluted human serum and ground-up mosquitoes, with the lowest detectable amount being 100 PFU\/mL. The GNRs described in this study can be used to enhance flavivirus diagnostic tests or to develop new, faster, and more accurate diagnostic techniques. Additionally, the functionalized GNRs presented here are promising for supporting virological surveillance studies in mosquitoes. Our findings highlight a fast and highly sensitive method for detecting Orthoflavivirus in both human and mosquito samples, with a detection limit as low as 100 PFU\/mL.<\/jats:p>","DOI":"10.3390\/s25154775","type":"journal-article","created":{"date-parts":[[2025,8,4]],"date-time":"2025-08-04T16:41:54Z","timestamp":1754325714000},"page":"4775","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Direct Detection of Orthoflavivirus via Gold Nanorod Plasmon Resonance"],"prefix":"10.3390","volume":"25","author":[{"given":"Erica Milena de Castro","family":"Ribeiro","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Federal University of Ouro Preto, Ouro Preto 35402-136, Brazil"},{"name":"Oswaldo Cruz Foundation, Ren\u00e9 Rachou Institute, FIOCRUZ, Belo Horizonte 30190-009, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruna de Paula","family":"Dias","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Federal University of Ouro Preto, Ouro Preto 35402-136, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9883-1392","authenticated-orcid":false,"given":"Cyntia Silva","family":"Ferreira","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Federal University of Ouro Preto, Ouro Preto 35402-136, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samara Mayra Soares Alves dos","family":"Santos","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Federal University of Ouro Preto, Ouro Preto 35402-136, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4030-0879","authenticated-orcid":false,"given":"Rajiv Gandhi","family":"Gopalsamy","sequence":"additional","affiliation":[{"name":"Division of Phytochemistry and Drug Design, Department of Biosciences, Rajagiri College of Social Sciences (Autonomous), Kochi 683104, Kerala, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Estef\u00e2nia Mara do Nascimento","family":"Martins","sequence":"additional","affiliation":[{"name":"Nuclear Technology Development Center, CDTN, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cintia Lopes de Brito","family":"Magalh\u00e3es","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Federal University of Ouro Preto, Ouro Preto 35402-136, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1416-8694","authenticated-orcid":false,"given":"Flavio Guimar\u00e3es da","family":"Fonseca","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4289-384X","authenticated-orcid":false,"given":"Luiz Felipe Leomil","family":"Coelho","sequence":"additional","affiliation":[{"name":"Vaccine Laboratory, Department of Microbiology and Immunology, Federal University of Alfenas, Alfenas 37130-001, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristiano","family":"Fantini","sequence":"additional","affiliation":[{"name":"Department of Physics, Federal University of Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luiz Orlando","family":"Ladeira","sequence":"additional","affiliation":[{"name":"Department of Physics, Federal University of Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1721-1547","authenticated-orcid":false,"given":"Lysandro Pinto","family":"Borges","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Federal University of Sergipe, S\u00e3o Crist\u00f3v\u00e3o 49400-000, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2472-8111","authenticated-orcid":false,"given":"Breno de Mello","family":"Silva","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Federal University of Ouro Preto, Ouro Preto 35402-136, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1099\/vir.0.036525-0","article-title":"Molecular Evolution of the Insect-specific Flaviviruses","volume":"93","author":"Cook","year":"2012","journal-title":"J. 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