适体
核酸
核糖核酸
多路复用
折叠(DSP实现)
环介导等温扩增
计算生物学
化学
核酸法
组合化学
计算机科学
纳米技术
分子生物学
生物
生物化学
DNA
材料科学
基因
工程类
电气工程
电信
作者
Zhaoqing Yan,Anli A. Tang,Amit Eshed,Zachary M. Ticktin,Soma Chaudhary,Duo Ma,Griffin McCutcheon,Yudan Li,Kaiyue Wu,Sanchari Saha,Jonathan Alcántar‐Fernández,José L Moreno-Camacho,Abraham Campos‐Romero,James J. Collins,Peng Yin,Alexander A. Green
出处
期刊:Cold Spring Harbor Laboratory - medRxiv
日期:2023-06-06
被引量:1
标识
DOI:10.1101/2023.06.02.23290873
摘要
Rapid, simple, and low-cost diagnostic technologies are crucial tools for combatting infectious disease. Here, we describe a class of aptamer-based RNA switches called aptaswitches that recognize specific target nucleic acid molecules and respond by initiating folding of a reporter aptamer. Aptaswitches can detect virtually any sequence and provide a fast and intense fluorescent readout, generating signals in as little as 5 minutes and enabling detection by eye with minimal equipment. We demonstrate that aptaswitches can be used to regulate folding of six different fluorescent aptamer/fluorogen pairs, providing a general means of controlling aptamer activity and an array of different reporter colors for multiplexing. By coupling isothermal amplification reactions with aptaswitches, we reach sensitivities down to 1 RNA copy/μL in one-pot reactions. Application of multiplexed one-pot reactions against RNA extracted from clinical saliva samples yields an overall accuracy of 96.67% for detection of SARS-CoV-2 in 30 minutes. Aptaswitches are thus versatile tools for nucleic acid detection that can be readily integrated into rapid diagnostic assays.
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