{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T06:34:42Z","timestamp":1788590082452,"version":"build-2803163510"},"reference-count":40,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2023,5,3]],"date-time":"2023-05-03T00:00:00Z","timestamp":1683072000000},"content-version":"vor","delay-in-days":2,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004001","name":"Guizhou Science and Technology Department","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2023,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Human mpox is a zoonotic disease, similar to smallpox, caused by the mpox virus, which is further subdivided into Congo Basin and West African clades with different pathogenicity. In this study, a novel diagnostic protocol utilizing clustered regularly interspaced short palindromic repeats (CRISPR)\/CRISPR\u2010associated 12a nuclease (CRISPR\/Cas12a)\u2010mediated recombinase polymerase amplification (RPA) was developed to identify mpox in the Congo Basin and West Africa (CRISPR\u2010RPA). Specific RPA primers targeting <jats:italic>D14L<\/jats:italic> and <jats:italic>ATI<\/jats:italic> were designed. CRISPR\u2010RPA assay was performed using various target templates. In the designed CRISPR\u2010RPA reaction system, the exponentially amplified RPA amplification products with a protospacer adjacent motif (PAM) site can locate the Cas12a\/crRNA complex to its target regions, which successfully activates the CRISPR\/Cas12a effector and achieves ultrafast <jats:italic>trans<\/jats:italic>\u2010cleavage of a single\u2010stranded DNA probe. The limit of detection for the CRISPR\u2010RPA assay was 10 copies per reaction for <jats:italic>D14L\u2010<\/jats:italic> and <jats:italic>ATI<\/jats:italic>\u2010plasmids. No cross\u2010reactivity was observed with non\u2010mpox strains, confirming the high specificity of the CRISPR\u2010RPA assay for distinguishing between the Congo Basin and West African mpox. The CRISPR\u2010RPA assay can be completed within 45\u2009min using real\u2010time fluorescence readout. Moreover, the cleavage results were visualized under UV light or an imaging system, eliminating the need for a specialized apparatus. In summary, the developed CRISPR\/RPA assay is a visual, rapid, sensitive, and highly specific detection technique that can be used as an attractive potential identification tool for Congo Basin and West African mpox in resource\u2010limited laboratories.<\/jats:p>","DOI":"10.1002\/jmv.28757","type":"journal-article","created":{"date-parts":[[2023,5,3]],"date-time":"2023-05-03T08:06:04Z","timestamp":1683101164000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Development of a CRISPR\/Cas12a\u2010recombinase polymerase amplification assay for visual and highly specific identification of the Congo Basin and West African strains of mpox virus"],"prefix":"10.1002","volume":"95","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4677-9470","authenticated-orcid":false,"given":"Xinggui","family":"Yang","sequence":"first","affiliation":[{"name":"Guizhou Provincial Center for Disease Control and Prevention Guiyang Guizhou P.R. 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