Fu Jinyu, Li Junjie, Chen Jing, Li Yabei, Liu Jiajia, Su Xin, Shi Shuobo
Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China.
Department of Neurosurgery, People's Hospital of Shijiazhuang, Shijiazhuang, China.
Crit Rev Biotechnol. 2022 Nov;42(7):1061-1078. doi: 10.1080/07388551.2021.1983757. Epub 2021 Oct 27.
Specific and sensitive detection of nucleic acids is essential to clinical diagnostics and biotechnological applications. Currently, amplification steps are necessary for most detection methods due to the low concentration of nucleic acid targets in real samples. Although amplification renders high sensitivity, poor specificity is prevalent because of the lack of highly accurate precise strategies, resulting in significant false positives and false negatives. Nucleases exhibit high catalytic activity for nucleic acid cleavage which is regulated in a programmable manner. This review focuses on the latest progress in nucleic acid testing methods based on the target-activated nucleases. It summarizes the property of enzymes such as CRISPR/Cas, Argonautes, and some gene-editing irrelevant nucleases, which have been leveraged to create highly specific and sensitive nucleic acid testing tools. We elaborate on recent advances in the field of nuclease-mediated DNA recognition techniques for nucleic acid detection, and discuss its future applications and challenges in molecular diagnostics.
核酸的特异性和灵敏检测对于临床诊断和生物技术应用至关重要。目前，由于实际样本中核酸靶标的浓度较低，大多数检测方法都需要扩增步骤。尽管扩增可带来高灵敏度，但由于缺乏高度准确精确的策略，普遍存在特异性差的问题，导致大量假阳性和假阴性。核酸酶对核酸切割表现出高催化活性，且这种活性以可编程的方式受到调控。本综述聚焦于基于靶标激活核酸酶的核酸检测方法的最新进展。它总结了诸如CRISPR/Cas、Argonautes等酶以及一些与基因编辑无关的核酸酶的特性，这些酶已被用于创建高度特异性和灵敏的核酸检测工具。我们详细阐述了核酸酶介导的用于核酸检测的DNA识别技术领域的最新进展，并讨论了其在分子诊断中的未来应用和挑战。
