Статья

Cas12aVDet: A CRISPR/Cas12a-Based Platform for Rapid and Visual Nucleic Acid Detection

Bei WangState Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, 200432, ChinaRui WangCollege of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, ChinaDaqi WangState Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, 200432, ChinaJian WuCollege of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, ChinaJixi LiState Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, 200432, ChinaJin WangCollege of Life Sciences, Shanghai Normal University, Shanghai 200234, ChinaHuihui LiuExperimental Center of Forestry in North China, Chinese Academy of Forestry, Beijing, 102300, ChinaYongming WangState Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, 200432, China

2019en

ABI

Аннотация

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.

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Идентификаторы

DOI: 10.1021/acs.analchem.9b01526

Цитирования и источники

Цитирований: 2Использованных источников: 0

Показатели — AkademScholar