通过集成CRISPR/ cas13a辅助系统检测CYP2C19基因

IF 2.6 3区 化学 Q2 CHEMISTRY, ANALYTICAL
Zhaokang Mai, Tao Zhou and Zhun Lin
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引用次数: 0

摘要

临床使用氯吡格雷时应考虑CYP2C19基因单核苷酸多态性(snp),对预测氯吡格雷治疗后出血和血栓形成风险具有重要的指导价值。由于其单核苷酸错配的特异性,CRISPR/Cas系统越来越多地用于SNP检测。同时检测多个snp对于快速鉴定CYP2C19基因型很重要,但目前还没有检测多种CYP2C19 snp的方法。本研究提出了一种新的集成系统,将三个CYP2C19基因的PCR反应和CRISPR/Cas检测集成在一个设备上,实现了快速、灵敏、特异的检测。在我们的设计中,用三种不同大小的磁珠捕获样品中的靶核酸,通过磁力拖动通过不同的区域,进行CYP2C19*2、CYP2C19*3和CYP2C19*17基因的PCR扩增反应和CRISPR/Cas13a检测。请注意,磁珠是通过不同孔径的微孔PC膜进行分类的。本研究具有广阔的临床应用前景,为临床氯吡格雷给药提供了有利的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Detecting CYP2C19 genes through an integrated CRISPR/Cas13a-assisted system

Detecting CYP2C19 genes through an integrated CRISPR/Cas13a-assisted system

CYP2C19 gene single nucleotide polymorphisms (SNPs) should be considered in the clinical use of clopidogrel as they have important guiding value for predicting the risk of bleeding and thrombosis after clopidogrel treatment. The CRISPR/Cas system is increasingly used for SNP detection owing to its single-nucleotide mismatch specificity. Simultaneous detection of multiple SNPs for rapid identification of the CYP2C19 genotype is important, but there is no method to detect a wide variety of CYP2C19 SNPs. This study proposes a new integrated system that integrates the PCR reaction and CRISPR/Cas detection of three CYP2C19 genes on a device, achieving rapid, sensitive, and specific detection. In our design, magnetic beads with three different sizes capture target nucleic acid from the sample, which are dragged through different areas by magnetic force, for PCR amplification reaction and CRISPR/Cas13a detection of CYP2C19*2, CYP2C19*3 and CYP2C19*17 genes. Note that magnetic beads were sorted via microporous PC membranes of different apertures. This study exhibits a broad clinical application prospect and provides a favorable tool for clinical clopidogrel administration.

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来源期刊
Analytical Methods
Analytical Methods CHEMISTRY, ANALYTICAL-FOOD SCIENCE & TECHNOLOGY
CiteScore
5.10
自引率
3.20%
发文量
569
审稿时长
1.8 months
期刊介绍: Early applied demonstrations of new analytical methods with clear societal impact
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