{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:43:45Z","timestamp":1792806225656,"version":"4.5.0"},"reference-count":23,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2023,6,30]],"date-time":"2023-06-30T00:00:00Z","timestamp":1688083200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2023,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The recent major worldwide outbreak of monkeypox virus (MPXV) has highlighted the urgent need for accurate MPXV detection methods. Although quantitative PCR\u00a0(qPCR) technique is currently the gold standard for MPXV diagnosis, the high costs associated with the technique and the need for complex instrumentation, limits its application in resource\u2010poor settings. CRISPR technology has developed rapidly in recent years and provides an effective tool for point\u2010of\u2010care testing\u00a0pathogen identification. Here, we exploited the cleavage properties of the Cas12a enzyme and Cas13a enzyme, to detect the MPXV specific genes, F3L gene and B6R gene, respectively. We developed two detection protocols: a 2\u2010step method in which the CRISPR Dual System reaction and the multiplex recombinase polymerase amplification\u00a0reaction were carried out in separate tubes and a single\u2010tube method in which both reactions were carried out in one tube. Evaluation of the two\u00a0methods showed that our protocol can detect the MPXV genome down to 10\u00b0 copies\/\u03bcL with good specificity and no cross\u2010reactivity with other poxviruses pseudoviruses, and bacteria. Mock positive samples were used to assess clinical applicability, with the results showing satisfactory concordance with the qPCR method for parallel testing. In conclusion, our study provides a reliable molecular diagnostic strategy for detection of MPXV.<\/jats:p>","DOI":"10.1002\/jmv.28905","type":"journal-article","created":{"date-parts":[[2023,6,30]],"date-time":"2023-06-30T07:31:39Z","timestamp":1688110299000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Creating an ultra\u2010sensitive detection platform for monkeypox virus DNA based on CRISPR technology"],"prefix":"10.1002","volume":"95","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5401-5983","authenticated-orcid":false,"given":"Tong","family":"Jiang","sequence":"first","affiliation":[{"name":"The First Affiliated Hospital of Anhui Medical University Hefei Anhui China"},{"name":"Anhui Public Health Clinical Center Hefei Anhui China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ge","family":"Li","sequence":"additional","affiliation":[{"name":"The First Affiliated Hospital of Anhui 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