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用于病原体检测的Cas14a1介导的核酸检测平台。

Cas14a1-mediated nucleic acid detectifon platform for pathogens.

作者信息

Ge Xiaolin, Meng Tian, Tan Xiao, Wei Yangdao, Tao Zhenzhen, Yang Zhiqing, Song Fengge, Wang Peng, Wan Yi

机构信息

State Key Laboratory of Marine Resource Utilization in South China Sea, Marine College, Key Laboratory of Tropical Biological Resources of Ministry of Education, School of Life and Pharmaceutical Sciences, Hainan University, 56 Renmin Road, Haikou, 570228, China.

Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing, 100084, China.

出版信息

Biosens Bioelectron. 2021 Oct 1;189:113350. doi: 10.1016/j.bios.2021.113350. Epub 2021 May 17.

Abstract

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease (Cas) based biosensing system provides a novel genomic diagnostic tool for pathogenic detection. However, most of the discovered Cas effectors have poor single strand DNA (ssDNA) target recognition capability with the constraint of protospacer adjacent motif (PAM) sites, which are not suitable for universal pathogenic diagnosis. Herein, we developed a highly sensitive and specific fluorescence tool for bacterial detection by utilizing the unique collateral cleavage activity of a Cas14a1-mediated nucleic acid detection platform (CMP). We combine CMP with molecular amplification to build a CRISPR-Cas based bioanalysis technique, offering fast nucleic acid detection with high sensitivity and specificity. This technique can identify different species of pathogens in milk samples with excellent accuracy. The CMP technique is a promising platform for pathogenic genomic diagnostic in biomedicine and food safety field.

摘要

基于成簇规律间隔短回文重复序列(CRISPR)相关核酸酶(Cas)的生物传感系统为病原体检测提供了一种新型的基因组诊断工具。然而,由于原间隔序列临近基序(PAM)位点的限制,大多数已发现的Cas效应蛋白对单链DNA(ssDNA)靶标的识别能力较差,不适用于通用的病原体诊断。在此,我们利用Cas14a1介导的核酸检测平台(CMP)独特的附带切割活性,开发了一种用于细菌检测的高灵敏度和特异性荧光工具。我们将CMP与分子扩增相结合,构建了一种基于CRISPR-Cas的生物分析技术,可实现高灵敏度和特异性的快速核酸检测。该技术能够以优异的准确性识别牛奶样本中的不同病原体种类。CMP技术是生物医学和食品安全领域用于病原体基因组诊断的一个有前景的平台。

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