Department of Biotechnology, College of Engineering, The University of Suwon, Hwaseong 18323, Korea.
Shandong Province Key Laboratory of Detection Technology of Tumor Markers, Linyi University, Linyi 276005, China.
Anal Methods. 2024 Oct 10;16(39):6599-6614. doi: 10.1039/d4ay01147c.
Currently, various infectious pathogens and bacterial toxins as well as heavy metal pollution pose severe threats to global environmental health and the socio-economic infrastructure. Therefore, there is a pressing need for rapid, sensitive, and convenient visual molecular detection methods. The rapidly evolving detection approach based on clustered regularly interspaced short palindromic repeats (CRISPR)/associated nucleases (Cas) has opened a new frontier in the field of molecular diagnostics. This paper reviews the development of visual detection methods in recent years based on different Cas and analyzes their advantages and disadvantages as well as the challenges of future research. Firstly, different CRISPR/Cas effectors and their working principles in the diagnosis of various diseases are briefly reviewed. Subsequently, the article focuses on the development of visual readout signals in point-of-care testing using laboratory-based CRISPR/Cas technology, including colorimetric, fluorescence, and lateral flow analysis. Finally, the challenges and prospects of visual detection methods based on CRISPR/Cas technology are discussed.
目前,各种传染性病原体和细菌毒素以及重金属污染对全球环境健康和社会经济基础设施构成了严重威胁。因此,迫切需要快速、灵敏、方便的可视化分子检测方法。基于成簇规律间隔短回文重复序列(CRISPR)/相关核酸酶(Cas)的快速发展的检测方法在分子诊断领域开辟了一个新的前沿。本文综述了近年来基于不同 Cas 的可视化检测方法的发展,并分析了它们的优缺点以及未来研究的挑战。首先,简要回顾了不同的 CRISPR/Cas 效应物及其在各种疾病诊断中的工作原理。随后,本文重点介绍了基于实验室的 CRISPR/Cas 技术在即时检测中的可视化读出信号的发展,包括比色法、荧光法和侧向流动分析。最后,讨论了基于 CRISPR/Cas 技术的可视化检测方法的挑战和前景。