Weihong Yin, Kai Hu, Bingwen Yu, Tao Zhang, Haohua Mei, Bowen Zhang, Zheyu Zou, Liping Xia, Yehong Gui, Juxing Yin, Wei Jin and Ying Mu
{"title":"使用微孔封闭和单氮化丙啶辅助的数字 CRISPR 微流控平台快速灵敏地检测大肠杆菌 O157:H7","authors":"Weihong Yin, Kai Hu, Bingwen Yu, Tao Zhang, Haohua Mei, Bowen Zhang, Zheyu Zou, Liping Xia, Yehong Gui, Juxing Yin, Wei Jin and Ying Mu","doi":"10.1039/D4LC00672K","DOIUrl":null,"url":null,"abstract":"<p >\r\n <em>Escherichia coli</em> O157:H7 is a major foodborne pathogen that poses a significant threat to food safety and human health. Rapid and sensitive detection of viable <em>Escherichia coli</em> O157:H7 can effectively prevent food poisoning. Here, we developed a microwell-confined and propidium monoazide-assisted digital CRISPR microfluidic platform for rapid and sensitive detection of viable <em>Escherichia coli</em> O157:H7 in food samples. The reaction time is significantly reduced by minimizing the microwell volume, yielding qualitative results in 5 min and absolute quantitative results in 15 min. With the assistance of propidium monoazide, this platform can eliminate the interference from 99% of dead <em>Escherichia coli</em> O157:H7. The direct lysis method obviates the need for a complex nucleic acid extraction process, offering a limit of detection of 3.6 × 10<small><sup>1</sup></small> CFU mL<small><sup>−1</sup></small> within 30 min. Our results demonstrated that the platform provides a powerful tool for rapid detection of <em>Escherichia coli</em> O157:H7 and provides reliable guidance for food safety testing.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":null,"pages":null},"PeriodicalIF":6.1000,"publicationDate":"2024-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Fast and sensitive detection of viable Escherichia coli O157:H7 using a microwell-confined and propidium monoazide-assisted digital CRISPR microfluidic platform†\",\"authors\":\"Weihong Yin, Kai Hu, Bingwen Yu, Tao Zhang, Haohua Mei, Bowen Zhang, Zheyu Zou, Liping Xia, Yehong Gui, Juxing Yin, Wei Jin and Ying Mu\",\"doi\":\"10.1039/D4LC00672K\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p >\\r\\n <em>Escherichia coli</em> O157:H7 is a major foodborne pathogen that poses a significant threat to food safety and human health. Rapid and sensitive detection of viable <em>Escherichia coli</em> O157:H7 can effectively prevent food poisoning. Here, we developed a microwell-confined and propidium monoazide-assisted digital CRISPR microfluidic platform for rapid and sensitive detection of viable <em>Escherichia coli</em> O157:H7 in food samples. The reaction time is significantly reduced by minimizing the microwell volume, yielding qualitative results in 5 min and absolute quantitative results in 15 min. With the assistance of propidium monoazide, this platform can eliminate the interference from 99% of dead <em>Escherichia coli</em> O157:H7. The direct lysis method obviates the need for a complex nucleic acid extraction process, offering a limit of detection of 3.6 × 10<small><sup>1</sup></small> CFU mL<small><sup>−1</sup></small> within 30 min. Our results demonstrated that the platform provides a powerful tool for rapid detection of <em>Escherichia coli</em> O157:H7 and provides reliable guidance for food safety testing.</p>\",\"PeriodicalId\":85,\"journal\":{\"name\":\"Lab on a Chip\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":6.1000,\"publicationDate\":\"2024-08-26\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Lab on a Chip\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://pubs.rsc.org/en/content/articlelanding/2024/lc/d4lc00672k\",\"RegionNum\":2,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Lab on a Chip","FirstCategoryId":"5","ListUrlMain":"https://pubs.rsc.org/en/content/articlelanding/2024/lc/d4lc00672k","RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
Fast and sensitive detection of viable Escherichia coli O157:H7 using a microwell-confined and propidium monoazide-assisted digital CRISPR microfluidic platform†
Escherichia coli O157:H7 is a major foodborne pathogen that poses a significant threat to food safety and human health. Rapid and sensitive detection of viable Escherichia coli O157:H7 can effectively prevent food poisoning. Here, we developed a microwell-confined and propidium monoazide-assisted digital CRISPR microfluidic platform for rapid and sensitive detection of viable Escherichia coli O157:H7 in food samples. The reaction time is significantly reduced by minimizing the microwell volume, yielding qualitative results in 5 min and absolute quantitative results in 15 min. With the assistance of propidium monoazide, this platform can eliminate the interference from 99% of dead Escherichia coli O157:H7. The direct lysis method obviates the need for a complex nucleic acid extraction process, offering a limit of detection of 3.6 × 101 CFU mL−1 within 30 min. Our results demonstrated that the platform provides a powerful tool for rapid detection of Escherichia coli O157:H7 and provides reliable guidance for food safety testing.
期刊介绍:
Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.