A universal and label-free aptasensor for fluorescent detection of ATP and thrombin based on SYBR Green I dye
A universal and label-free aptasensor for fluorescent detection of ATP and thrombin based on SYBR Green I dye
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基于 SYBR Green I 染料的通用、无标记适体传感器，用于 ATP 和凝血酶的荧光检测
DOI：
10.1016/j.bios.2012.10.064
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发表时间：
2013-04-15
影响因子：
12.6
通讯作者：
Chai, Yaqin
中科院分区：
文献类型：
--
作者：
Kong, Ling;Xu, Jin;Chai, Yaqin
利用SYBR绿色I（SGI）染料，建立了一种简便、通用的基于核酸适体的无标记荧光检测方法，用于蛋白质和生物小分子的选择性和灵敏度的检测。这种强大的多功能生物传感策略依赖于SGI的荧光关闭变化，这是由靶诱导的适体结构转换引起的。在与靶标结合后，适体从各自的cDNA/适体双链体解离，导致嵌入dsDNA的SGI释放到溶液中并淬灭相应的荧光强度。这种靶诱导的构象变化和从DNA双链体释放适体基本上导致SGI的荧光信号的变化，从而构成了我们的基于适体的无标记荧光生物传感器用于特定靶分析的机制。在优化的条件下，我们的方法表现出高灵敏度和选择性的ATP和凝血酶的定量与低检测限（23.4 nM和1.1 nM，分别）。与以前报道的基于适配体的ATP和凝血酶检测方法相比，这种无标记方法具有选择性，简单，方便和成本效益高，无需对探针或靶标进行任何化学标记。因此，本发明的策略可以容易地应用于靶向广泛的生物分子的生物传感器。(C)2012爱思唯尔有限公司版权所有。
A facile and universal aptamer-based label-free approach for selective and sensitive fluorescence detection of proteins and small biomolecules by using the SYBR Green I (SGI) dye is developed. This robust versatile biosensing strategy relies on fluorescence turn-off changes of SGI, resulting from target-induced structure switching of aptamers. Upon binding with the targets, the aptamers dissociate from the respective cDNA/aptamer duplexes, leading to the release of the dsDNA-intercalated SGI into solution and the quenching of the corresponding fluorescence intensities. Such target-induced conformational changes and release of aptamers from the DNA duplexes essentially lead to the change in the fluorescence signal of the SGI and thus constitute the mechanism of our aptamer-based label-free fluorescence biosensor for specific target analyses. Under optimized conditions, our method exhibits high sensitivity and selectivity for the quantification of ATP and thrombin with low detection limits (23.4 nM and 1.1 nM, respectively). Compared with previous reported methods for aptamer-based detection of ATP and thrombin, this label-free approach is selective, simple, convenient and cost-efficient without any chemical labeling of the probe or the target. Therefore, the present strategy could be easily applicable to biosensors that target a wide range of biomolecules. (C) 2012 Elsevier B.V. All rights reserved.