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基于 CRISPR/Cas 系统的现场检测进展。

Advances in Field Detection Based on CRISPR/Cas System.

机构信息

Department of Epidemiology, College of Public Health, Zhengzhou University, No. 100 Kexue Avenue, Zhengzhou, Henan Province 450000, China.

出版信息

ACS Synth Biol. 2021 Nov 19;10(11):2824-2832. doi: 10.1021/acssynbio.1c00401. Epub 2021 Oct 29.

Abstract

Rapid and accurate diagnostic methods are essential to interrupt outbreaks of infectious diseases such as COVID-19. However, the most commonly used nucleic acid detection method, qPCR or RT-qPCR, takes several hours to complete and requires highly sophisticated equipment. Recently, an emerging nucleic acid detection method based on the CRISPR/Cas system has reduced the reliance on qPCR. It has several important features that make it suitable for on-site POCT (point-of-care testing), including short detection cycles, low cost, high sensitivity, and the ability to be combined with different readout methods. This review briefly introduces the steps of CRISPR/Cas detection and then summarizes the current advances of CRISPR/Cas-based POCT from four steps: nucleic acid extraction, target amplification, CRISPR/Cas-based signal generation, and signal output. Finally, we discuss the advantages and challenges of CRISPR-based POCT and describe the future research perspectives for CRISPR.

摘要

快速准确的诊断方法对于中断 COVID-19 等传染病的爆发至关重要。然而,最常用的核酸检测方法 qPCR 或 RT-qPCR 需要数小时才能完成,并且需要高度复杂的设备。最近,一种基于 CRISPR/Cas 系统的新兴核酸检测方法减少了对 qPCR 的依赖。它具有几个重要的特点,使其适合现场 POCT(即时护理测试),包括检测周期短、成本低、灵敏度高以及能够与不同的读出方法相结合。这篇综述简要介绍了 CRISPR/Cas 检测的步骤,然后从四个步骤总结了基于 CRISPR/Cas 的 POCT 的最新进展:核酸提取、靶扩增、基于 CRISPR/Cas 的信号生成和信号输出。最后,我们讨论了基于 CRISPR 的 POCT 的优势和挑战,并描述了 CRISPR 的未来研究前景。

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