{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T11:03:13Z","timestamp":1790593393050,"version":"4.1.0"},"reference-count":36,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T00:00:00Z","timestamp":1788652800000},"content-version":"vor","delay-in-days":5,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2021YFF0703600"],"award-info":[{"award-number":["2021YFF0703600"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbial Biotechnology"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>In 2024, Monkeypox virus (MPXV) clade I has triggered outbreaks in several countries world\u2010wide. MPXV clade I demonstrates enhanced virulence and transmissibility, with a case fatality rate reaching 10%. In response, we have developed a one\u2010pot detection assay specifically targeting MPXV clade I, combining recombinase polymerase amplification (RPA) and the CRISPR\/Cas12a system. The assay can be completed within 40\u2009min and achieved a 95% limit of detection (LOD95) of 27.16\u2009copies\/\u03bcL. No cross\u2010reactivity was observed with MPXV clade II or other tested viral templates, including Vaccinia virus (Tiantan strain). A preliminary room\u2010temperature evaluation showed that the assay retained detectable performance at 25\u00b0C, supporting its potential use in equipment\u2010limited settings. The assay also showed good intra\u2010assay and inter\u2010assay repeatability for recombinant plasmid templates, with all coefficient of variation (CV) values below 10%. In simulated clinical samples, the RPA\u2010CRISPR\/Cas12a assay detected more low\u2010concentration plasmid\u2010spiked samples than quantitative polymerase chain reaction (qPCR). These results indicate that the established assay is specific, sensitive, repeatable, and easy to perform, providing a practical tool for field\u2010based screening and decentralized detection of MPXV clade I.<\/jats:p>","DOI":"10.1111\/1751-7915.70437","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T10:27:31Z","timestamp":1788776851000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["One\u2010Pot\n                    <scp>RPA<\/scp>\n                    \u2010\n                    <scp>CRISPR<\/scp>\n                    \/Cas12a Assay With Visual Readout for the Ultra\u2010Specific Detection of Monkeypox Virus Clade I"],"prefix":"10.1111","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-9534-2086","authenticated-orcid":false,"given":"Boyi","family":"Li","sequence":"first","affiliation":[{"name":"State Key Laboratory for Diagnosis and Treatment of Severe 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Jilin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-1244-3745","authenticated-orcid":false,"given":"Rong","family":"Gao","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine Jilin University  Jilin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-1203-4859","authenticated-orcid":false,"given":"Huanxin","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine Jilin University  Jilin 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