Yan Zhaoqing, Eshed Amit, Tang Anli A, Arevalos Nery R, Ticktin Zachary M, Chaudhary Soma, Ma Duo, McCutcheon Griffin, Li Yudan, Wu Kaiyue, Saha Sanchari, Alcantar-Fernandez Jonathan, Moreno-Camacho Jose L, Campos-Romero Abraham, Collins James J, Yin Peng, Green Alexander A
Department of Biomedical Engineering, Boston University, Boston, MA, USA.
Molecular Biology, Cell Biology & Biochemistry Program, Graduate School of Arts and Sciences, Boston University, Boston, MA, USA.
Chem. 2024 Jul 11;10(7):2220-2244. doi: 10.1016/j.chempr.2024.03.015. Epub 2024 Apr 12.
Rapid, simple, and low-cost diagnostic technologies are crucial tools for combatting infectious disease. We describe a class of aptamer-based RNA switches or aptaswitches that recognize target nucleic acid molecules and initiate folding of a reporter aptamer. Aptaswitches can detect virtually any sequence and provide an intense fluorescent readout without intervening enzymes, generating signals in as little as 5 minutes and enabling detection by eye with minimal equipment. Aptaswitches can be used to regulate folding of seven fluorogenic aptamers, providing a general means of controlling aptamers and an array of multiplexable reporter colors. Coupling isothermal amplification reactions with aptaswitches, we reach sensitivities down to 1 RNA copy/μL in one-pot reactions. Application of multiplexed all-in-one reactions against RNA 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 are readily integrated into rapid diagnostic assays.
快速、简单且低成本的诊断技术是对抗传染病的关键工具。我们描述了一类基于适配体的RNA开关或适配开关，它们能够识别靶核酸分子并启动报告适配体的折叠。适配开关几乎可以检测任何序列，无需中间酶即可提供强烈的荧光读数，只需5分钟即可产生信号，并且使用最少的设备就能肉眼检测。适配开关可用于调节七种荧光适配体的折叠，提供了一种控制适配体的通用方法以及一系列可复用的报告颜色。将等温扩增反应与适配开关相结合，我们在一锅反应中实现了低至1个RNA拷贝/微升的灵敏度。针对临床唾液样本中的RNA进行多重一体化反应，在30分钟内检测SARS-CoV-2的总体准确率达到96.67%。因此，适配开关是用于核酸检测的多功能工具，很容易集成到快速诊断检测中。
