Yang Hao, Ledesma-Amaro Rodrigo, Gao Hong, Ren Yao, Deng Ruijie
College of Biomass Science and Engineering, Healthy Food Evaluation Research Center, Sichuan University, Chengdu, 610065, China.
Department of Bioengineering, Imperial College Centre for Synthetic Biology, Imperial College London, London, SW7 2AZ, UK.
Biosens Bioelectron. 2023 May 15;228:115189. doi: 10.1016/j.bios.2023.115189. Epub 2023 Mar 2.
Pathogenic biosafety is a worldwide concern. Tools for analyzing pathogenic biosafety, that are precise, rapid and field-deployable, are highly demanded. Recently developed biotechnological tools, especially those utilizing CRISPR/Cas systems which can couple with nanotechnologies, have enormous potential to achieve point-of-care (POC) testing for pathogen infection. In this review, we first introduce the working principle of class II CRISPR/Cas system for detecting nucleic acid and non-nucleic acid biomarkers, and highlight the molecular assays that leverage CRISPR technologies for POC detection. We summarize the application of CRISPR tools in detecting pathogens, including pathogenic bacteria, viruses, fungi and parasites and their variants, and highlight the profiling of pathogens' genotypes or phenotypes, such as the viability, and drug-resistance. In addition, we discuss the challenges and opportunities of CRISPR-based biosensors in pathogenic biosafety analysis.
病原生物安全是全球关注的问题。人们迫切需要精确、快速且可现场部署的病原生物安全分析工具。最近开发的生物技术工具，尤其是那些利用可与纳米技术相结合的CRISPR/Cas系统的工具，在实现病原体感染的即时检测（POC）方面具有巨大潜力。在本综述中，我们首先介绍用于检测核酸和非核酸生物标志物的II类CRISPR/Cas系统的工作原理，并重点介绍利用CRISPR技术进行即时检测的分子检测方法。我们总结了CRISPR工具在检测病原体（包括致病细菌、病毒、真菌和寄生虫及其变体）中的应用，并重点介绍了病原体基因型或表型（如活力和耐药性）的分析。此外，我们还讨论了基于CRISPR的生物传感器在病原生物安全分析中的挑战与机遇。
