摘要
病原菌的快速准确检测是实现疫情高效防控、疾病精准治疗、污染环境及时处置的关键。而现有的病原菌现场快速检测技术,主要以定性分析为主,假阳性/假阴性受到诟病,检测准确性仍有待提升,亟待发展基于新原理、新方法的病原菌快速检测技术。基于CRISPR(clustered regularly interspaced short palindromic repeats)的生物传感技术因具有高灵活性(对不同的基因靶点只需改变crRNA序列)、高特异性(单碱基分辨)、高灵敏(优于10^(-18)mol/L浓度)、可编程、可模块化、低成本、可在各种体外介质中高效稳定运行等独特优势,打破了传统分子诊断与检测技术的局限性,正在成为下一代病原菌检测技术的引领者。在该技术中,Cas效应蛋白被用作高特异性的序列识别元件,结合不同的生物传感机制,即可用于病原菌的高特异性快速灵敏检测。在总结CRISPR/Cas生物传感技术原理的基础上,综述了用于病原菌检测的CRISPR/Cas12和CRISPR/Cas13生物传感技术研究进展。通过阐述CRISPR/Cas生物传感技术在实际应用中面临的挑战,展望其未来的发展前景。
Rapid and accurate detection of pathogens is essential to achieve efficient epidemic prevention and control,accurate treatment of diseases,and timely disposal of polluted environment.However,the existing on-site rapid detection methods of pathogenic bacteria mainly focuses on qualitative analysis.False positive/negative results exist and their detection accuracy still needs to be improved.It is urgent to develop rapid detection technologies of pathogenic bacteria by taking use of new principles and methods.The CRISPR(clustered regularly interspaced short palindromic repeats)based biosensors have several unique advantages,such as high flexibility(only needing to change the crRNA sequence for different target genes),high specificity(single base resolution),high sensitivity(better than 10^(-18) mol/L concentration),programmability,modularity,low cost,and high efficiency and stability in various in vitro media.It has become the leader of the next generation of pathogen detection technologies without the limitations of traditional molecular diagnosis and detection technologies.In this technology,Cas effector proteins are used as highly specific sequence recognition elements.Through combined with various biosensor mechanisms,they can be used for rapid and sensitive detection of pathogens with high specificity.After summarizing the principle of the CRISPR/Cas biosensor technology,the research progress of the CRISPR/Cas12 and CRISPR/Cas13 biosensor technologies for pathogen detection was reviewed.The challenges of the CRISPR/Cas biosensor technology in practical application are discussed,and its future developments are prospected.
作者
徐文娟
宋丹
陈丹
龙辉
陈禹保
龙峰
XU Wen-juan;SONG Dan;CHEN Dan;LONG Hui;CHEN Yu-bao;LONG Feng(School of Environmental&Natural Resources,Renmin University,Beijing 100875,China;Institute of Product Quality Inspection,Guangxi Zhuang Autonomous Region,Nanning 530007,China;Institute of Laboratory Animal Sciences,Beijing 100021,China)
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2021年第8期67-74,共8页
China Biotechnology
基金
国家自然科学基金(21077063)资助项目。
