{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T05:54:53Z","timestamp":1781848493509,"version":"3.54.5"},"reference-count":48,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T00:00:00Z","timestamp":1781827200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>\n                    <jats:italic>Enterovirus A71<\/jats:italic>\n                    (EVA71) is a major pathogen responsible for hand, foot, and mouth disease (HFMD) outbreaks and poses a considerable threat to public health. In this work, we established a molecular detection system that integrates reverse transcription multiple cross displacement amplification (RT-MCDA) with CRISPR\/Cas12a technology, targeting the VP1 gene region for EVA71 identification. This platform, termed EVA71-RT-MCDA-CRISPR, utilizes RT-MCDA to achieve efficient pre-amplification of the target sequence, followed by sequence-specific recognition mediated by CRISPR\/Cas12a. The detection results can be observed both through real-time fluorescence analysis and visual inspection under blue light illumination. The assay exhibits outstanding sensitivity, achieving a limit of detection as low as 3.36 copies per reaction, while retaining high specificity against non-EVA71 templates. Validation using clinical specimens further confirmed the diagnostic reliability of this RT-MCDA-CRISPR platform. The successful establishment of the EVA71-RT-MCDA-CRISPR system demonstrates its potential as a rapid, user-friendly, highly sensitive and specific method for on-site EVA71 detection, serving as a valuable tool for clinical diagnosis and epidemiological surveillance.\n                  <\/jats:p>","DOI":"10.3389\/fmicb.2026.1883920","type":"journal-article","created":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T05:47:59Z","timestamp":1781848079000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["A CRISPR\/Cas12a-based reverse transcription multiple cross displacement amplification technique for sensitive, rapid, and highly specific detection of human enterovirus A71 in clinical application"],"prefix":"10.3389","volume":"17","author":[{"given":"Xujian","family":"Zhang","sequence":"first","affiliation":[{"name":"School of Basic Medical Sciences, Guizhou Key Laboratory of Microbio and Infectious Disease Prevention and Control, Guizhou Medical University","place":["Guiyang, Guizhou, China"]},{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Ye","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]},{"name":"Department of Basic Clinical Laboratory Medicine, School of Clinical Laboratory Science, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]},{"name":"Department of Basic Clinical Laboratory Medicine, School of Clinical Laboratory Science, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yumei","family":"Cao","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]},{"name":"School of Public Health, Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mao","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]},{"name":"Department of Basic Clinical Laboratory Medicine, School of Clinical Laboratory Science, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Wei","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yonglin","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Yuan","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guo","family":"Guo","sequence":"additional","affiliation":[{"name":"School of Basic Medical Sciences, Guizhou Key Laboratory of Microbio and Infectious Disease Prevention and Control, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, The First People's Hospital of Guiyang","place":["Guiyang, Guizhou, China"]},{"name":"Department of Basic Clinical Laboratory Medicine, School of Clinical Laboratory Science, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,6,19]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"198129","DOI":"10.1016\/j.virusres.2020.198129","article-title":"iSCAN: an RT-LAMP-coupled CRISPR-Cas12 module for rapid, sensitive detection of SARS-CoV-2","volume":"288","author":"Ali","year":"2020","journal-title":"Virus 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