{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T16:57:45Z","timestamp":1781197065617,"version":"3.54.1"},"reference-count":21,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T00:00:00Z","timestamp":1761868800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Open Project Program of Chongqing Key Laboratory of Highly Pathogenic Microbes","award":["2025ZDSYS005"],"award-info":[{"award-number":["2025ZDSYS005"]}]},{"name":"National Key Research and Development Program of China","award":["2025YFC2609603"],"award-info":[{"award-number":["2025YFC2609603"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Diagnosis of dengue virus infections typically relies on RT-PCR-based methods, for which reliable positive controls are essential. Viral RNA is an ideal positive control, but its inherent instability poses a major challenge. Herein, we report a simple and effective method for stabilizing dengue virus RNA by immobilizing it onto silk fibroin films (RNA-SFFs). We evaluated various substrate surfaces for RNA-SFFs preparation and found that the inner surface of sealable bags is optimal for uniform film formation and easy harvesting. Screening different silk fibroin concentrations revealed that even low concentrations (2.8%) effectively preserved RNA well and kept Ct constant for up to 16 days at 25 \u00b0C, 37 \u00b0C, and even 45 \u00b0C (extreme weather for transportations). Due to its rapid film formation and ease of peeling, 7% silk fibroin was selected. Notably, the RNA-SFFs demonstrated robust resistance to UV irradiation, with no significant Ct value changes after 4 h of exposure. Long-term stability testing at \u221220 \u00b0C, 25 \u00b0C, and 37 \u00b0C showed that dengue serotype 1\u20134 RNA-SFFs remained stable for the entire duration of the study\u2014up to 56 weeks (approximately 14 months)\u2014at all tested temperatures. These results demonstrate that RNA-SFFs are highly stable, portable, and practical as positive controls for dengue diagnostics, with strong potential for use in on-site and resource-limited settings.<\/jats:p>","DOI":"10.3390\/v17111452","type":"journal-article","created":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T13:47:58Z","timestamp":1762177678000},"page":"1452","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Long-Term Stabilization of Dengue Virus RNA at 37 \u00b0C for 14 Months Using Silk Fibroin Films"],"prefix":"10.3390","volume":"17","author":[{"given":"Nuo","family":"Wang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Virology and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"He","sequence":"additional","affiliation":[{"name":"Chongqing Key Laboratory of Highly Pathogenic Microbes, Chongqing Disease Prevention and Public Health Research Center Construction Program, Chongqing Center for Disease Control and Prevention (Chongqing Academy of Preventive Medicine), Chongging 400707, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bohan","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Virology and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"},{"name":"University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9948-8880","authenticated-orcid":false,"given":"Hongping","family":"Wei","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Virology and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"},{"name":"University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9024-0793","authenticated-orcid":false,"given":"Junping","family":"Yu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Virology and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"},{"name":"University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1007\/s00705-025-06235-3","article-title":"Dengue: Epidemiology, diagnosis methods, treatment options, and prevention strategies","volume":"170","author":"Kothari","year":"2025","journal-title":"Arch. 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