文献类型：期刊文献

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

作者：Wang, Bei[1] Wang, Rui[2] Wang, Daqi[1] Wu, Jian[2] Li, Jixi[1] Wang, Jin[4] Liu, Huihui[3] Wang, Yongming[1]

第一作者：Wang, Bei

通信作者：Wang, YM[1]

机构：[1]Fudan Univ, Zhongshan Hosp, Sch Life Sci, State Key Lab Genet Engn, Shanghai 200432, Peoples R China;[2]Zhejiang Univ, Coll Biosyst Engn & Food Sci, Hangzhou 310058, Zhejiang, Peoples R China;[3]Chinese Acad Forestry, Expt Ctr Forestry North China, Beijing 102300, Peoples R China;[4]Shanghai Normal Univ, Coll Life Sci, Shanghai 200234, Peoples R China

年份：2019

卷号：91

期号：19

起止页码：12156-12161

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20194207534985);Scopus(收录号：2-s2.0-85072790655);WOS:【SCI-EXPANDED(收录号:WOS:000488993000007)】；

基金：This work was supported by grants from the National Natural Science Foundation of China (Grant 81870199), the National Basic Research Program of China (Grant 2015CB943300), the Foundation for Innovative Research Group of the National Natural Science Foundation of China (Grant 31521003), and Thousand Youth Talents Plan Project of Thousand Youth Talents. We thank Dr. S.-G. Ong for revision of the manuscript.

语种：英文

外文关键词：Biomolecules - Chemical detection - Fluorophores - Nucleic acids

摘要：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.