{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T20:09:02Z","timestamp":1792613342173,"version":"4.5.0"},"reference-count":28,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,6,6]],"date-time":"2026-06-06T00:00:00Z","timestamp":1780704000000},"content-version":"vor","delay-in-days":5,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T00:00:00Z","timestamp":1780272000000},"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":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Co\u2010infection with both hepatitis B virus (HBV) and hepatitis D virus (HDV) can aggravate the severity of the end\u2010stage liver disease and accelerate its progression. However, no combined diagnostic method for HDV and HBV nucleic acids exists. In this study, we have developed a highly sensitive and specific dual detection method for HDV RNA and HBV DNA using the CRISPR\u2010Cas system. We established a dual detection method combining CRISPR\u2010Cas12a with recombinase polymerase amplification (RAA) for HBV, and CRISPR\u2010Cas13a with RT\u2010RAA for HDV. Validation was performed using specimens from 70 co\u2010infected patients. RAA primers and crRNAs were designed and optimized to establish a dual fluorescence detection method (DF) and lateral flow strip\u2010based dual detection (DL) within the same CRISPR\u2010Cas13a\/Cas12a system for HDV RNA and HBV DNA. The system demonstrated 100% specificity, and both DF and DL methods exhibited a sensitivity of 10\u2009copies\/\u03bcL for synthetic positive plasmids and samples. Peak fluorescence detection was achieved with T7 RNA polymerase, while the best detection efficiency was at ssRNA: ssDNA ratio of 1:1.5. In the validation of plasma samples from 70 co\u2010infected clinical patients, the positive concordance rates for RT\u2010RAA\u2010CRISPR\u2010Cas13a\/Cas12a DF and DL were 85.7% (60\/70) and 82.9% (58\/70), respectively. We developed a CRISPR\u2010Cas13a\/Cas12a\u2010based dual assay for sensitive, specific, and accurate detection of HDV RNA and HBV DNA, offering an effective tool for the early detection, treatment, and monitoring of HDV and HBV infections.<\/jats:p>","DOI":"10.1002\/jmv.70976","type":"journal-article","created":{"date-parts":[[2026,6,6]],"date-time":"2026-06-06T08:56:21Z","timestamp":1780736181000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["CRISPR\u2010Cas13a\/Cas12a Assisted Dual Portable and Visualized HDV and HBV Detection"],"prefix":"10.1002","volume":"98","author":[{"given":"Yuan","family":"Tian","sequence":"first","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing PR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaling","family":"Cao","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Shanghai General Hospital Shanghai Jiao Tong University School of Medicine Shanghai PR 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 PR 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 PR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zihao","family":"Fan","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing PR 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 PR 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 PR 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 PR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology Beijing PR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shengtao","family":"Zeng","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases Xinhua Hospital of Ili Kazakh Autonomous Prefecture Xinjiang PR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Ren","sequence":"additional","affiliation":[{"name":"Beijing Institute of Hepatology\/Beijing Youan Hospital Capital Medical University Beijing PR China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,6,6]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41575-023-00760-9"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.3350\/cmh.2014.20.3.223"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(22)01468-4"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2021.03.025"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2021.01.014"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2009.01.052"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2018-316601"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2020.04.008"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiz633"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.21042"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v12.i45.7365"},{"issue":"6","key":"e_1_2_10_13_1","first-page":"417","article-title":"Expression and Purification of Recombinant Hepatitis Delta Virus (HDV) Antigen for Use in a Diagnostic ELISA for HDV Infection Using the High\u2010Density Fermentation Strategy in Escherichia coli","volume":"29","author":"Lu X. X.","year":"2016","journal-title":"Biomedical and Environmental Sciences: BES"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2015.08.009"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2013.07.033"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.14309\/ctg.0000000000000509"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aam9321"},{"key":"e_1_2_10_18_1","unstructured":"I. M.LobatoandC. K.O'SullivanRecombinase Polymerase Amplification: Basics Applications and Recent Advances."},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar6245"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1039\/D2AY01536F"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2023.2177088"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2023.2276337"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.3390\/v9040075"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.3390\/v13122371"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jare.2019.03.009"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaq0179"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11427-021-2028-x"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1039\/D2AY01536F"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.3390\/v12050538"}],"container-title":["Journal of Medical Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.70976","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jmv.70976","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.70976","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T09:18:31Z","timestamp":1782465511000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmv.70976"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6]]},"references-count":28,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026,6]]}},"alternative-id":["10.1002\/jmv.70976"],"URL":"https:\/\/doi.org\/10.1002\/jmv.70976","archive":["Portico"],"relation":{},"ISSN":["0146-6615","1096-9071"],"issn-type":[{"value":"0146-6615","type":"print"},{"value":"1096-9071","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6]]},"assertion":[{"value":"2025-08-14","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-13","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-06","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70976"}}