{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T16:14:02Z","timestamp":1791994442781,"version":"4.4.0"},"reference-count":50,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T00:00:00Z","timestamp":1775692800000},"content-version":"vor","delay-in-days":8,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T00:00:00Z","timestamp":1775001600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Human metapneumovirus (HMPV) is a significant pathogen, causing widespread acute respiratory infections (ARIs). Rapid and accurate detection is crucial for timely diagnosis and outbreak control. To address the limitations of real\u2010time reverse transcription quantitative polymerase chain reaction (RT\u2010qPCR)\u2010based HMPV detection, we developed a novel method integrating recombinase\u2010aided amplification (RAA) with CRISPR\u2010Cas13a technology. Based on the alignment of 335 HMPV whole\u2010genome sequences, a conserved region of the nucleocapsid (\n                    <jats:italic>N<\/jats:italic>\n                    ) gene was selected for RAA primer and CRISPR RNA (crRNA) design. The concentrations of Cas protein, crRNA, and probe were optimized for specificity, sensitivity, and repeatability. A dual\u2010mode platform was established, combining a fluorescence assay for quantitative detection and a lateral flow assay (LFA) for visual, on\u2010site readout. Both methods were validated using 106 clinical samples and compared with RT\u2010qPCR. Both methods demonstrated high specificity (no cross\u2010reactivity with other respiratory viruses), a detection limit of 1\u2009copy\/\u03bcL, and excellent repeatability. Clinical validation showed complete concordance (\n                    <jats:italic>\u03ba<\/jats:italic>\n                    \u2009=\u20091,\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.001) between the fluorescence assay and RT\u2010qPCR, and high agreement for LFA (\n                    <jats:italic>\u03ba<\/jats:italic>\n                    \u2009=\u20090.924,\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.001), with a sensitivity of 94.23%, specificity of 98.15%, positive predictive value (PPV) of 98%, and negative predictive value (NPV) of 94.6%. This study presents an RAA\u2010CRISPR\/Cas13a dual\u2010mode platform for HMPV detection, providing a robust molecular diagnostic framework that paves the way for the future development of rapid testing assays.\n                  <\/jats:p>","DOI":"10.1002\/jmv.70921","type":"journal-article","created":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T11:12:18Z","timestamp":1775733138000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Dual\u2010Mode CRISPR\/Cas13a Assay for the Detection of Human Metapneumovirus in Clinical Respiratory Samples"],"prefix":"10.1002","volume":"98","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-1714-7784","authenticated-orcid":false,"given":"Shiyuan","family":"Zheng","sequence":"first","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"},{"name":"NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-8225-3275","authenticated-orcid":false,"given":"Chao","family":"Wang","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"},{"name":"NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0754-5383","authenticated-orcid":false,"given":"Yiman","family":"Huang","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"},{"name":"NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-4703-0251","authenticated-orcid":false,"given":"Xiaocao","family":"Meng","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"},{"name":"NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-3841-5323","authenticated-orcid":false,"given":"Dong","family":"Zhao","sequence":"additional","affiliation":[{"name":"Shijiazhuang Center for Disease Control and Prevention Shijiazhuang China"},{"name":"Hebei Key Laboratory of Intractable Pathogens Shijiazhuang China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-8969-4334","authenticated-orcid":false,"given":"Enrui","family":"Guan","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"},{"name":"NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-6435-0851","authenticated-orcid":false,"given":"Fenlian","family":"Ma","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"},{"name":"NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5337-9745","authenticated-orcid":false,"given":"Aijun","family":"Chen","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"},{"name":"NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8780-6191","authenticated-orcid":false,"given":"Lishu","family":"Zheng","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing China"},{"name":"NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and Prevention, China CDC Beijing 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