分析化学2025，Vol.53Issue(12)：1964-1978,中插1-中插4,19.DOI:10.19756/j.issn.0253-3820.251239
规律成簇间隔短回文重复序列及其相关蛋白系统用于无扩增核酸检测病原体的研究进展
Advances in Clustered Regularly Interspaced Short Palindromic Repeats-associated Proteins Systems for Amplification-Free Nucleic Acid Detection of Pathogen
摘要
Abstract
Accurate and rapid detection of pathogens is crucial for public health management.Although traditional nucleic acid detection techniques such as quantitative polymerase chain reaction(qPCR)and isothermal amplification offer high sensitivity,they face challenges such as complex operations and aerosol contamination,making them unsuitable for pathogen detection in point-of-care testing(POCT).In recent years,clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPR associated proteins(CRISPR/Cas)amplification-free nucleic acid detection,with its simplicity,specificity,and rapidity,are becoming new research area for pathogen detection in POCT.This paper reviewed the advances in CRISPR/Cas amplification-free nucleic acid detection systems in recent years.Five common Cas systems(Cas9,Cas12a,Cas12b,Cas13a and Cas12f)and their detection principles were introduced.Key amplification-free strategies were categorized,including ultrasensitive signals(including electrochemical,electrochemiluminescence,and novel optical methods)and various signal amplification systems(including crRNA/enzyme cascades and micro-confinement).The solutions for improving sample processing,operational simplicity,multiplex detection,and PAM flexibility were discussed,aiming to guide the development of next-generation platforms with high sensitivity,multiplexing,and robust POCT capability.Finally,the potential of CRISPR/Cas amplification-free detection for pathogen diagnosis were outlined,and key challenges for translation research including sensor stability,reagent standardization,and clinical validation were pointed out.关键词
规律成簇间隔短回文重复序列及其相关蛋白/病原体/分子诊断/核酸检测/生物传感器/评述Key words
Clustered regularly interspaced short palindromic repeats-associated proteins/Pathogen/Molecular diagnostics/Nucleic acid detection/Biosensors/Review引用本文复制引用
胡欣言,张倬毓,张鹏飞..规律成簇间隔短回文重复序列及其相关蛋白系统用于无扩增核酸检测病原体的研究进展[J].分析化学,2025,53(12):1964-1978,中插1-中插4,19.基金项目
同济大学医学+X交叉研究项目(No.2025-0650-YB-12)和上海市科委科技项目(No.22S31902000)资助. Supported by the Tongji University"Medical+X"Cross-Disciplinary Research Project(No.2025-0650-YB-12)and the Shanghai Municipal Science and Technology Commission(No.22S31902000). （No.2025-0650-YB-12）
