A Propidium Monoazide-Assisted Digital CRISPR/Cas12a Assay for Selective Detection of Live Bacteria in Sample.

IF 6.7 1区化学 Q1 CHEMISTRY, ANALYTICAL

Analytical Chemistry Pub Date : 2024-11-12 Epub Date: 2024-10-31 DOI:10.1021/acs.analchem.4c02204

Weihong Yin, Kai Hu, Yunxing Yang, Jianjian Zhuang, Zheyu Zou, Yuanjie Suo, Liping Xia, Jiale Li, Yehong Gui, Haohua Mei, Juxin Yin, Tao Zhang, Ying Mu

{"title":"A Propidium Monoazide-Assisted Digital CRISPR/Cas12a Assay for Selective Detection of Live Bacteria in Sample.","authors":"Weihong Yin, Kai Hu, Yunxing Yang, Jianjian Zhuang, Zheyu Zou, Yuanjie Suo, Liping Xia, Jiale Li, Yehong Gui, Haohua Mei, Juxin Yin, Tao Zhang, Ying Mu","doi":"10.1021/acs.analchem.4c02204","DOIUrl":null,"url":null,"abstract":"Escherichia coli O157:H7 (E. coli O157:H7) is a prominent pathogenic bacterium that poses serious risks to food safety and public health. Rapid and accurate detection of live E. coli O157:H7 is of great importance in food quality monitoring and clinical diagnosis. Here, we report a propidium monoazide-assisted nonamplification digital CRISPR/Cas12a assay for sensitive and rapid detection of live E. coli O157:H7. The incorporation of propidium monoazide into the method enables the selective detection of live bacteria by eliminating 98% of interference from the dead bacterial nucleic acid. Implemented on microfluidic digital chips, this method can achieve absolute quantification of nonamplified nucleic acid. The entire detection process of live bacteria can be completed within 120 min without the need for establishing a standard curve, and the sensitivity of the method reaches 1.2 × 103 CFU/mL. The method was validated using various samples, yielding results consistent with the plate counting method (Pearson's r = 0.9490). Consequently, this method holds significant potential for applications in fields requiring live bacterial detection.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":" ","pages":"17941-17949"},"PeriodicalIF":6.7000,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical Chemistry","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1021/acs.analchem.4c02204","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/10/31 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}

引用次数: 0

Abstract

Escherichia coli O157:H7 (E. coli O157:H7) is a prominent pathogenic bacterium that poses serious risks to food safety and public health. Rapid and accurate detection of live E. coli O157:H7 is of great importance in food quality monitoring and clinical diagnosis. Here, we report a propidium monoazide-assisted nonamplification digital CRISPR/Cas12a assay for sensitive and rapid detection of live E. coli O157:H7. The incorporation of propidium monoazide into the method enables the selective detection of live bacteria by eliminating 98% of interference from the dead bacterial nucleic acid. Implemented on microfluidic digital chips, this method can achieve absolute quantification of nonamplified nucleic acid. The entire detection process of live bacteria can be completed within 120 min without the need for establishing a standard curve, and the sensitivity of the method reaches 1.2 × 10³ CFU/mL. The method was validated using various samples, yielding results consistent with the plate counting method (Pearson's r = 0.9490). Consequently, this method holds significant potential for applications in fields requiring live bacterial detection.

Abstract Image

查看原文本刊更多论文

一种用于选择性检测样品中活细菌的单氮化丙啶辅助数字 CRISPR/Cas12a 检测法。

大肠杆菌 O157:H7（E. coli O157:H7）是一种突出的致病菌，对食品安全和公众健康构成严重威胁。快速准确地检测活大肠杆菌 O157:H7 对食品质量监测和临床诊断具有重要意义。在此，我们报告了一种丙啶单氮辅助的非扩增数字 CRISPR/Cas12a 检测方法，用于灵敏快速地检测活大肠杆菌 O157:H7。在该方法中加入单氮化丙啶可消除 98% 的死细菌核酸干扰，从而实现对活细菌的选择性检测。该方法在微流控数字芯片上实施，可实现对非扩增核酸的绝对定量。整个活菌检测过程可在 120 分钟内完成，无需建立标准曲线，灵敏度达到 1.2 × 103 CFU/mL。该方法使用各种样品进行了验证，结果与平板计数法一致（Pearson's r = 0.9490）。因此，该方法在需要活细菌检测的领域具有很大的应用潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Analytical Chemistry 化学-分析化学

CiteScore

12.10

自引率

12.20%

发文量

1949

审稿时长

1.4 months

期刊介绍： Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.