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通过 LAMP-CRISPR/Cas12b 与热不稳定尿嘧啶-DNA 糖基化酶的结合,实现对 的快速可视化检测,从而消除携带污染。

Rapid visual detection of by combining LAMP-CRISPR/Cas12b with heat-labile uracil-DNA glycosylase to eliminate carry-over contamination.

机构信息

Jiangsu Key Laboratory of Marine Biological Resources and Environment, Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Jiangsu Ocean University, Lianyungang 222005, China.

Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Ocean University, Lianyungang 222005, China.

出版信息

J Zhejiang Univ Sci B. 2023 Aug 15;24(8):749-754. doi: 10.1631/jzus.B2200705.

DOI:10.1631/jzus.B2200705
PMID:37551560
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10423967/
Abstract

is a major pathogen frequently found in seafood. Rapid and accurate detection of this pathogen is important for the control of bacterial foodborne diseases and to ensure food safety. In this study, we established a one-pot system that combines uracil-DNA glycosylase (UDG), loop-mediated isothermal amplification (LAMP), and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 12b (Cas12b) for detecting in seafood. This detection system can effectively perform identification using a single tube and avoid the risk of carry-over contamination.

摘要

是一种常见于海鲜的主要病原体。快速准确地检测这种病原体对于控制细菌性食源性疾病和确保食品安全非常重要。在本研究中,我们建立了一种一管式系统,该系统结合了尿嘧啶-DNA 糖基化酶(UDG)、环介导等温扩增(LAMP)和簇状规则间隔短回文重复序列(CRISPR)/CRISPR 相关蛋白 12b(Cas12b),用于检测海鲜中的 。该检测系统可以使用单个管有效地进行鉴定,避免了交叉污染的风险。

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本文引用的文献

1
Recent advances in the use of the CRISPR-Cas system for the detection of infectious pathogens.CRISPR-Cas 系统在检测传染性病原体中的最新应用进展。
J Zhejiang Univ Sci B. 2022 Nov 15;23(11):881-898. doi: 10.1631/jzus.B2200068.
2
CE-RAA-CRISPR Assay: A Rapid and Sensitive Method for Detecting in Seafood.CE-RAA-CRISPR检测法:一种用于海鲜检测的快速灵敏方法。
Foods. 2022 Jun 8;11(12):1681. doi: 10.3390/foods11121681.
3
Fast and sensitive detection of SARS-CoV-2 RNA using suboptimal protospacer adjacent motifs for Cas12a.使用非最优的 Cas12a 邻近基序快速灵敏地检测 SARS-CoV-2 RNA。
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Universally Stable and Precise CRISPR-LAMP Detection Platform for Precise Multiple Respiratory Tract Virus Diagnosis Including Mutant SARS-CoV-2 Spike N501Y.用于精准呼吸道多病毒诊断的通用稳定且精确的 CRISPR-LAMP 检测平台,包括突变型 SARS-CoV-2 刺突 N501Y。
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An improved recombinase polymerase amplification assay for visual detection of Vibrio parahaemolyticus with lateral flow strips.一种改良的重组酶聚合酶扩增检测法,用于通过侧流条带进行可视检测副溶血性弧菌。
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HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation.HOLMESv2:一种用于核酸检测和DNA甲基化定量的CRISPR-Cas12b辅助平台。
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CDetection: CRISPR-Cas12b-based DNA detection with sub-attomolar sensitivity and single-base specificity.CDetection:基于 CRISPR-Cas12b 的 DNA 检测,具有亚皮摩尔灵敏度和单碱基特异性。
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