摘要
作为当前最先进的检测方法,基于特定核苷酸序列的核酸检测法被广泛地应用于基因分型、病原体和遗传疾病诊断及环境监控等领域.CRISPR/Cas系统可在指导RNA的导引下精准高效地识别和切割目标DNA/RNA,被用于研发新一代高灵敏、高特异性的体外核酸检测技术.重点综述了 CRISPR/Cas9、CRISPR/Cas12、CRISPR/Cas13、CRISPR/Cas14 系统应用于体外核酸检测的作用机制及高通量多通道检测、目标核酸的富集与扩增和信号输出方式等信号传感方向的技术进展,介绍了 CRISPR/Cas核酸检测在监控内源性疾病、病毒、细菌和寄生虫中的新应用,讨论了其优缺点与面临的挑战,并展望了其未来的研究方向,为CRISPR/Cas核酸检测的转化提供参考和依据.
Abstract
As the most advanced detection method,the sequence-based nucleic acid detection is widely used in genotyping,the diagnosis of pathogens and genetic diseases,and environmental monitoring.The CRISPR/Cas system can accurately and efficiently identify and cleave target DNA/RNA under the guidance of guide RNA,and thus is adopted to develop the new generation of in vitro nucleic acid detection with a high sensitivity and specificity.Along with the advances in multiple CRISPR/Cas systems,more and more new CRISPR/Cas-based techniques and platforms have been established for pathogen nucleic acid detection and early disease diagnosis.This article reviews the action mechanism of CRISPR/Cas9,12,13,14 systems in nucleic acid detection,and the novel signal sensing technologies of CRISPR/Cas-based nucleic acid detection,including the high-throughput and multi-channel assays,the enrichment and amplification of target nucleic acids,and the methods of signal output.It further illustrates the new application of CRISPR/Cas nucleic acid test in monitoring endogenous diseases,virus,bacteria,and parasites,discusses its advantages,disadvantages,and challenges,and outlines its future research directions,providing the reference and basis for the translation of CRISPR/Cas-based nucleic acid detection.
维普
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