基于CRISPR/Cas系统诊断平台的研究进展

Research progress of diagnosis platforms based on CRISPR/Cas system

目前核酸检测技术已被广泛应用于临床实验室诊断,常规检测技术如实时荧光定量PCR技术耗时长且依赖特定的仪器设备。成簇规律间隔短回文重复序列(CRISPR)/CRISPR相关蛋白(Cas)系统是细菌和古细菌在与病毒斗争过程中获得的适应性免疫防御机制,已被发展成强大的基因组编辑技术。最近CRISPR领域的先驱团队基于Cas13a、Cas12a和新发现的Cas14蛋白开发出SHERLOCK、DETECTR等新型核酸检测工具,在传染性疾病的快速诊断、癌症中基因突变的检测和基因分型等方面意义重大,其灵敏度高、特异性强且快速经济,在临床分子诊断领域具有巨大潜力。本文综述了CRISPR/Cas系统的作用机制及新型诊断平台的原理和应用进展,总结了新型检测技术的优缺点并对其发展前景进行展望。

At present,nucleic acid testing technology has been widely used in clinical laboratory diagnosis.Conventional detection technique such as real-time PCR is complicated,time consuming,and dependent on specific instruments.Clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR associated protein(Cas)system is an adaptive immune defense system against viruses in bacteria and archaea,which has been developed into a powerful technology for genome editing.Recently,the leading groups engaged in CRISPR have set up new tools for nucleic acid detection based on Cas13a,Cas12a and newly discovered protein-Cas14,which plays an important role in rapid diagnosis of infectious diseases,detection of gene mutations in cancer and genotyping.Since they are ultrasensitive,specific,rapid and cost-effective,it is expected to bring great potential for molecular diagnosis.In this review,the mechanism of CRISPR/Cas system and the principle,the applications of the newly-developed diagnostic platforms are introduced.What′s more,the advantages,limitations and prospects of the technologies are summarized.