基于CRISPR/Cas系统的核酸即时检测技术现状及研究进展

Current status and progress of nucleic acid point-of-care testing technology based on CRISPR/Cas system

在全球疫情背景下,现场条件下的核酸检测在疫情防控中发挥越来越重要的作用,这促使核酸即时检测(point-ofcare testing,POCT)工具的开发面临空前的需求。基于规律间隔成簇短回文重复序列及其相关蛋白系统(clustered regularly interspaced short palindromic repeats-associated nuclease,CRISPR/Cas)应用于核酸POCT在近年来得到飞速发展并取得良好成果。在疫情封控区、海关以及灾区等特殊现场,可以将恒温放大技术与CRISPR有机结合并集成到小型手持设备中,这些技术使用起来更简单,非专业人员能够更快捷准确地进行感染源的核酸检测。这篇综述首先简要介绍了CRISPR/Cas系统和不同的效应蛋白,回顾了基于CRISPR/Cas系统特异识别和切割特性在核酸POCT中的不同应用策略,最后讨论CRISPR/Cas用于核酸POCT面临的挑战及未来的研究前景。

In the context of the global epidemic,nucleic acid detection under on-site conditions plays a more and more important role in epidemic prevention and control,which makes the development of nucleic acid point-of-care testing(POCT)tools facing unprecedented demand. The application of clustered regularly interspaced palindromic repeats-associated nuclease(CRISPR/Cas)to nucleic acid POCT has been developed rapidly and achieved good results in recent years. In the epidemic control areas,customs and disaster areas and other special sites,combining isothermal amplification techniques with CRISPR and being integrated into small handheld devices,these technologies are easier to use and non-professionals can quickly and accurately detect the nucleic acid source of infection. This review first briefly introduces the CRISPR/Cas system and different effector proteins,reviews the different application strategies in nucleic acid POCT based on the specific recognition and cleavage characteristics of CRISPR/Cas system,and finally discusses the challenges and future research prospects of CRISPR/Cas in nucleic acid POCT.