分析化学2024，Vol.52Issue(2)：187-197,11.DOI:10.19756/j.issn.0253-3820.231382
基于CRISPR/Cas技术的传染性病原精准便携化检测系统
A Precise and Portable Detection System for Infectious Pathogens Based on CRISPR/Cas Technology
摘要
Abstract
Nucleic acid-based molecular diagnostic methods are considered the gold standard for detecting infectious pathogens.However,when applied to portable or on-site rapid diagnostics,they still face various limitations and challenges,such as poor specificity,cumbersome operation,and portability difficulties.The CRISPR(Clustered regularly interspaced short palindromic repeats)/CRISPR-associated protein(Cas)-fluorescence detection method holds the potential to significantly enhance the specificity and signal-to-noise ratio of nucleic acid detection.In this study,we developed a portable grayscale reader detection system based on loop-mediated isothermal amplification(LAMP)-CRISPR/Cas.On one hand,in the presence of CRISPR RNA(crRNA),the CRISPR/Cas12a system was employed to achieve precise fluorescent detection of self-designed LAMP amplification reactions for influenza A and influenza B viruses.This further validated the high selectivity and versatility of the CRISPR/Cas system.On the other hand,the accompanying independently developed portable grayscale reader allowed for low-cost collection of fluorescence signals and high-reliability visual interpretation.At the end of the detection process,it directly provided positive or negative results.Practical sample analyses using this detection system have verified its reliability and utility,demonstrating that this system can achieve highly sensitive and highly specific portable analysis of influenza viruses.关键词
等温扩增/CRISPR-Cas/荧光/便携化检测/流行性病原/灰度读取仪Key words
Isothermal amplification/CRISPR-Cas/Fluorescence/Portable detection/Infectious pathogens/Grayscale reader引用本文复制引用
刘一辰,赵茹健,吕佰阳,宋德锋,唐艺丹,姜艳芳,李冰凌..基于CRISPR/Cas技术的传染性病原精准便携化检测系统[J].分析化学,2024,52(2):187-197,11.基金项目
国家自然科学基金项目(No.22004118)和吉林省科技发展计划重点研发项目(No.20230203193SF)资助.Supported by the National Natural Science Foundation of China(No.22004118)and the Key Research and Development Program of Science and Technology Development Plan of Jilin Province,China(No.20230203193SF). （No.22004118）
