{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T16:39:34Z","timestamp":1788194374204,"version":"build-2803163510"},"reference-count":44,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T00:00:00Z","timestamp":1764633600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research and Development Program of China","award":["2024YFC2607800"],"award-info":[{"award-number":["2024YFC2607800"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Dengue fever has become a major global public health challenge due to its rapidly in-creasing incidence. Rapid on-site detection of dengue virus (DENV) is critical for early diagnosis, timely patient isolation, and outbreak control. In this study, dengue virus serotype 2 (DENV-2), the predominant strain circulating in tropical and subtropical regions, was selected as the target pathogen. We established a one-tube rapid detection assay that integrates the HUDSON nucleic acid extraction-free protocol, reverse transcription recombinase-aided amplification (RT-RAA), and CRISPR\/Cas12a-mediated trans cleavage activity. The method achieved a detection limit of 1 \u00d7 102 copies\/\u03bcL for simulated infected samples and exhibited no cross-reactivity with other DENV serotypes (DENV-1, DENV-3, DENV-4) or with other arboviruses, including Zika, Japanese encephalitis, yellow fever, and chikungunya viruses. The assay demonstrated high sensitivity and specificity across various sample types, including mosquitoes, rodents, blood, and cultured cells, with results consistent with quantitative PCR (qPCR). Requiring only basic equipment such as a water bath, the system enables on-site detection of DENV-2 within 1 h. This simple, cost-effective, and reliable assay provides a practical tool for field-based DENV-2 surveillance and supports effective public health responses in resource-limited settings.<\/jats:p>","DOI":"10.3390\/v17121579","type":"journal-article","created":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T12:39:43Z","timestamp":1764679183000},"page":"1579","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Development and Application of a Rapid Field Detection Technology for DENV-2 Based on the HUDSON Nucleic Acid Extraction-Free\/RT-RAA\/CRISPR-Cas12a System"],"prefix":"10.3390","volume":"17","author":[{"given":"Chang","family":"Tan","sequence":"first","affiliation":[{"name":"Characteristic Key Laboratory of Modern Pathogen Biology, School of Basic Medicine Sciences, Guizhou Medical University, Guian New Area, Guiyang 561113, China"},{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siyu","family":"Xing","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoxue","family":"Xie","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8047-2473","authenticated-orcid":false,"given":"Xiaoli","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohui","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenhao","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lifang","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Zhou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiahong","family":"Wu","sequence":"additional","affiliation":[{"name":"Characteristic Key Laboratory of Modern Pathogen Biology, School of Basic Medicine Sciences, Guizhou Medical University, Guian New Area, Guiyang 561113, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunxiao","family":"Li","sequence":"additional","affiliation":[{"name":"Characteristic Key Laboratory of Modern Pathogen Biology, School of Basic Medicine Sciences, Guizhou Medical University, Guian New Area, Guiyang 561113, China"},{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1016\/j.jmii.2020.03.007","article-title":"Dengue hemorrhagic fever-A systemic literature review of current perspectives on pathogenesis, prevention and control","volume":"53","author":"Wang","year":"2020","journal-title":"J. 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