Ultrasensitive detection of microRNA with isothermal amplification and a time-resolved fluorescence sensor
Ultrasensitive detection of microRNA with isothermal amplification and a time-resolved fluorescence sensor
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DOI：
10.1016/j.bios.2014.01.058
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发表时间：
2014-07-15
影响因子：
12.6
通讯作者：
Zhou, Fanfan
中科院分区：
文献类型：
--
作者：
Wang, Ke;Zhang, Kai;Zhou, Fanfan
MicroRNAs(MiRNAs)在众多生物和细胞过程中发挥着重要作用。MiRNAs的水平可以作为细胞事件或疾病诊断的有用生物标志物，因此，敏感和选择性地检测miRNAs的方法对于miRNA的发现、研究和临床诊断是必不可少的。在这里，我们开发了一种新的方法，通过两步指数放大反应(EXPAR)和一个时间分辨荧光传感器在实际样品中定量检测miRNA的表达水平，低至对大气分子的敏感性。该方法具有较高的灵敏度，在纯条件下，miRNA的检出限为0.1am。该方法对区分miRNA家族成员之间的差异具有很高的选择性，从而为定量检测miRNA提供了一种有前途的替代方法。皇冠版权所有(C)2014由Elsevier B.V.出版。保留所有权利。
MicroRNAs (miRNAs) play important roles in a plethora of biological and cellular processes. The levels of miRNAs can be useful biomarkers for cellular events or disease diagnosis, thus the method for sensitive and selective detection of miRNAs is imperative to miRNA discovery, study, and clinical diagnosis. Here we develop a novel method to quantify miRNA expression levels as low as attomolar sensitivity by twostage exponential amplification reaction (EXPAR) and a time-resolved fluorescence sensor in real samples. The method reveals superior sensitivity with a detection limit of miRNA of 0.1 aM under pure conditions. The method also shows the high selectivity for discriminating differences between miRNA family members, thus providing a promising alternative to standard approaches for quantitative detection of miRNA. Crown Copyright (c) 2014 Published by Elsevier B.V. All rights reserved.