{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T02:07:01Z","timestamp":1787623621845,"version":"build-2736575974"},"reference-count":36,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2024,12,20]],"date-time":"2024-12-20T00:00:00Z","timestamp":1734652800000},"content-version":"vor","delay-in-days":19,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002799","name":"Capital Medical University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002799","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":[[2024,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Hepatitis E virus (HEV) poses a serious threat to both public health and animal food safety, thereby highlighting the demands for rapid, sensitive, and easy\u2010to\u2010use detection. This study aimed to develop a One\u2010Pot assay using CRISPR\/Cas13a for detecting HEV RNA, suitable for point\u2010of\u2010care testing (POCT) in resource\u2010limited settings. CRISPR\/Cas13a combined with reverse transcription polymerase chain reaction (RT\u2010PCR) and reverse transcription recombinase\u2010aided amplification (RT\u2010RAA) was applied to a One\u2010Pot assay device. Additionally, a large cohort of HEV\u2010infected patient (154) and animal (104) specimens was utilized for validation. The RT\u2010PCR\/RT\u2010RAA\u2009+\u2009CRISPR\/Cas13a assays for HEV RNA detection (genotypes: HEV\u20101, HEV\u20103, and HEV\u20104) were established, optimized, and validated, achieving a limit of detection (LoD) of 1\u2009copy\/\u03bcL and 100% specificity. In the application validation for HEV infection, the positive rates of the RT\u2010PCR\u2009+\u2009CRISPR and RT\u2010RAA\u2009+\u2009CRISPR assays were 98.6% and 89.6% for patients, and 96.6% and 88.8% for animals, respectively, which were superior to those of RT\u2010qPCR. Furthermore, sample rapid lysis, reagent lyophilization, and the One\u2010Pot device were integrated to construct a One\u2010Pot assay with an LoD of 10\n                    <jats:sup>2<\/jats:sup>\n                    \u2009copies\/\u03bcL. Despite slight decreases in sensitivity, the One\u2010Pot assay significantly reduces the assay time to 35\u2009min, making it easy to perform, minimizing contamination, and meeting the requirements for screening. We developed a One\u2010Pot assay of HEV RNA using the CRISPR\/Cas13a which effectively realizes a POCT test and maximizes the impetus for POCT implementation and shows potential as a valuable tool for detecting and monitoring HEV infection.\n                  <\/jats:p>","DOI":"10.1002\/jmv.70115","type":"journal-article","created":{"date-parts":[[2024,12,20]],"date-time":"2024-12-20T06:09:02Z","timestamp":1734674942000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["One\u2010Pot Assay Based on CRISPR\/Cas13a Technology for HEV RNA Point\u2010of\u2010Care Testing"],"prefix":"10.1002","volume":"96","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0011-8394","authenticated-orcid":false,"given":"Zihao","family":"Fan","sequence":"first","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ling","family":"Xu","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaling","family":"Cao","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tianxu","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Infectious Disease Center, School of Basic Medical Sciences Peking University Health Science Center Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan","family":"Tian","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenzhen","family":"Pan","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinkang","family":"Mo","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Wang","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xianru","family":"Zhu","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yao","family":"Gao","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangying","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Calvin Q.","family":"Pan","sequence":"additional","affiliation":[{"name":"NYU Langone Medical Center New York University School of Medicine New York City New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Microbiology 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