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Our previous study showed that a simple and highly repeatable plaque assay could be used for enterovirus 71 (EV\u2010A71). Now, we show that using a subclone of a clinical EV\u2010A71 isolate and a rhabdomyosarcoma cell line (RD), a plaque assay based on an EV\u2010A71\/RD model could exhibit the most rapid formation of plaques (&lt;2 days), with much higher repeatability and consistency. Inspired by a plaque inhibitory test for testing ribavirin and interferon, as well as a plaque reduction neutralization test, this modified method has been used to establish a convenient system by using 96\u2010well plates for screening anti\u2010EV\u2010A71 drugs from a 130\u2010compound library containing multiple types of inhibitors. Nine candidate effective compounds for EV\u2010A71 have been screened out, and among them, nobiletin (<jats:italic>flavonoid<\/jats:italic>) was found to be a novel effective compound at the concentration of 10\u2009\u03bcM. Our findings imply that this improved method based on an EV\u2010A71\/RD model proved to be a potential high\u2010throughput method in screening novel antiviral drugs for EV\u2010A71. Undoubtedly, this method can also be applied to other viruses that can produce an obvious cytopathic effect.<\/jats:p>","DOI":"10.1002\/jmv.25463","type":"journal-article","created":{"date-parts":[[2019,3,22]],"date-time":"2019-03-22T10:28:38Z","timestamp":1553250518000},"page":"1440-1447","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["An antiviral drug screening system for enterovirus 71 based on an improved plaque assay: A potential high\u2010throughput method"],"prefix":"10.1002","volume":"91","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8685-4378","authenticated-orcid":false,"given":"Yingxian","family":"Yin","sequence":"first","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases, Guangdong Institute of Gastroenterology The Sixth Affiliated Hospital, Sun Yat\u2010Sen University Guangzhou Guangdong China"},{"name":"Department of Clinical Laboratory The Sixth 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