Cas12aVDet: A CRISPR/Cas12a-Based Platform for Rapid and Visual Nucleic Acid Detection
Cas12aVDet: A CRISPR/Cas12a-Based Platform for Rapid and Visual Nucleic Acid Detection
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Cas12aVDet：基于 CRISPR/Cas12a 的快速、可视化核酸检测平台
DOI：
10.1021/acs.analchem.9b01526
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发表时间：
2019-10-01
影响因子：
7.4
通讯作者：
Wang, Yongming
中科院分区：
文献类型：
--
作者：
Wang, Bei;Wang, Rui;Wang, Yongming
一种快速、灵敏的核酸检测方法至关重要。最近，基于rna引导的CRISPR/Cas12a核酸酶的方法在核酸检测方面显示出很大的前景。然而，在目前的方法中，DNA扩增和随后的Cas12a切割是分开的，整个过程需要长达2小时。最重要的是，打开盖子的操作增加了气溶胶污染的风险。在本研究中，我们提出了一种基于CRISPR/ cas12的快速核酸检测方法，命名为“Cas12aVDet”。将重组酶聚合酶扩增（recombinase polymerase amplification， RPA）与Cas12a裂解结合在一个反应体系中，可在30分钟内完成检测，避免脱帽污染。检测信号可以在蓝光下用肉眼观察到。该方法可在单分子水平上检测DNA，支原体污染检测准确率100%，具有广泛的核酸检测应用前景。
A rapid and sensitive method is crucial for nucleic acid detection. Recently, RNA-guided CRISPR/Cas12a nuclease-based methods present great promise for nucleic acid detection. In the present methods, however, DNA amplification and subsequent Cas12a cleavage is separated and the whole process takes as long as 2 h. Most importantly, the uncapping operation increases the risk of aerosol contamination. In this study, we propose a CRISPR/Cas12a-based method named "Cas12aVDet" for rapid nucleic acid detection. By integrating recombinase polymerase amplification (RPA) with Cas12a cleavage in a single reaction system, the detection can be accomplished in 30 min and uncapping contamination can be avoided. The detection signal can be observed by the naked eye under blue light. This method could detect DNA at single molecule level and demonstrated 100% accuracy for mycoplasma contamination detection, presenting great potential for a variety of nucleic acid detection applications.