核酸
核酸检测
清脆的
多路复用
DNA
环介导等温扩增
计算生物学
核糖核酸
化学
分子生物学
纳米技术
生物
计算机科学
生物化学
材料科学
基因
电信
作者
Jonathan S. Gootenberg,Omar O. Abudayyeh,Max J. Kellner,Julia Joung,James J. Collins,Feng Zhang
出处
期刊:Science
[American Association for the Advancement of Science (AAAS)]
日期:2018-04-27
卷期号:360 (6387): 439-444
被引量:1686
标识
DOI:10.1126/science.aaq0179
摘要
Rapid detection of nucleic acids is integral for clinical diagnostics and biotechnological applications. We recently developed a platform termed SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) that combines isothermal preamplification with Cas13 to detect single molecules of RNA or DNA. Through characterization of CRISPR enzymology and application development, we report here four advances integrated into SHERLOCK version 2 (SHERLOCKv2) (i) four-channel single-reaction multiplexing with orthogonal CRISPR enzymes; (ii) quantitative measurement of input as low as 2 attomolar; (iii) 3.5-fold increase in signal sensitivity by combining Cas13 with Csm6, an auxiliary CRISPR-associated enzyme; and (iv) lateral-flow readout. SHERLOCKv2 can detect Dengue or Zika virus single-stranded RNA as well as mutations in patient liquid biopsy samples via lateral flow, highlighting its potential as a multiplexable, portable, rapid, and quantitative detection platform of nucleic acids.
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