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The emerging clustered regularly interspaced short palindromic repeats (CRISPR)\/Cas has been regarded as a promising point\u2010of\u2010care (POC) strategy for nucleic acids detection. However, how to achieve CRISPR\/Cas multiplex biosensing remains a challenge. Here, an affordable means termed CRISPR\u2010RDB (CRISPR\u2010based reverse dot blot) for multiplex target detection in parallel, which possesses the advantages of high sensitivity and specificity, cost\u2010effectiveness, instrument\u2010free, ease to use, and visualization is reported. CRISPR\u2010RDB integrates the trans\u2010cleavage activity of CRISPR\u2010Cas12a with a commercial RDB technique. It utilizes different Cas12a\u2010crRNA complexes to separately identify multiple targets in one sample and converts targeted information into colorimetric signals on a piece of accessible nylon membrane that attaches corresponding specific\u2010oligonucleotide probes. It has demonstrated that the versatility of CRISPR\u2010RDB by constructing a four\u2010channel system to simultaneously detect influenza A, influenza B, respiratory syncytial virus, and SARS\u2010CoV\u20102. With a simple modification of crRNAs, the CRISPR\u2010RDB can be modified to detect human papillomavirus, saving two\u2010thirds of the time compared to a commercial PCR\u2010RDB kit. Further, a user\u2010friendly microchip system for convenient use, as well as a smartphone app for signal interpretation, is engineered. CRISPR\u2010RDB represents a desirable option for multiplexed biosensing and on\u2010site diagnosis.<\/jats:p>","DOI":"10.1002\/advs.202204689","type":"journal-article","created":{"date-parts":[[2022,11,28]],"date-time":"2022-11-28T20:59:55Z","timestamp":1669669195000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":57,"title":["CRISPR\/Cas12a\u2010Enabled Multiplex Biosensing Strategy Via an Affordable and Visual Nylon Membrane Readout"],"prefix":"10.1002","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3448-6515","authenticated-orcid":false,"given":"Tao","family":"Hu","sequence":"first","affiliation":[{"name":"The Children's Hospital Zhejiang University School of Medicine National Clinical Research Center for Child Health  Hangzhou Zhejiang 310052 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinxin","family":"Ke","sequence":"additional","affiliation":[{"name":"The Children's Hospital Zhejiang University School of Medicine National Clinical Research Center for Child Health  Hangzhou Zhejiang 310052 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Li","sequence":"additional","affiliation":[{"name":"The Children's Hospital Zhejiang University School of Medicine National Clinical Research Center for Child Health  Hangzhou Zhejiang 310052 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Lin","sequence":"additional","affiliation":[{"name":"International Peace Maternity &amp; Child Health Hospital Shanghai Municipal Key Clinical Specialty Institute of Embryo\u2010Fetal Original Adult Disease School of Medicine Shanghai Jiao Tong University  Shanghai 200030 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ajuan","family":"Liang","sequence":"additional","affiliation":[{"name":"Center of Reproductive Medicine Shanghai First Maternity and Infant Hospital Tongji University School of Medicine  Shanghai 201204 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yangjing","family":"Ou","sequence":"additional","affiliation":[{"name":"International Peace Maternity &amp; Child Health Hospital Shanghai Municipal Key Clinical Specialty Institute of Embryo\u2010Fetal Original Adult Disease School of Medicine Shanghai Jiao Tong University  Shanghai 200030 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuanxia","family":"Chen","sequence":"additional","affiliation":[{"name":"School of Materials Science and Engineering University of Jinan  Jinan Shandong 250022 China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,11,28]]},"reference":[{"key":"e_1_2_8_1_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2110672119"},{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.7b04551"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41551-021-00755-4"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1039\/D0CS00609B"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-22664-5"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.3390\/diagnostics11050823"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.02965-20"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2020.112867"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1261\/rna.076232.120"},{"key":"e_1_2_8_10_1","doi-asserted-by":"crossref","first-page":"e0179121","DOI":"10.1128\/jvi.01791-21","volume":"96","author":"Huang Y.","year":"2022","journal-title":"J. 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