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基于适体构象转变的 8-羟基-2'-脱氧鸟苷的超灵敏无标记检测法。

An ultrasensitive label-free assay of 8-hydroxy-2'-deoxyguanosine based on the conformational switching of aptamer.

机构信息

College of Public Health, University of South China, Hengyang 421001, PR China; College of Public Health, Dali University, Dali 671000, PR China.

College of Public Health, University of South China, Hengyang 421001, PR China.

出版信息

Biosens Bioelectron. 2014 Aug 15;58:22-6. doi: 10.1016/j.bios.2014.02.047. Epub 2014 Feb 26.

Abstract

We developed a highly sensitive label-free assay of 8-hydroxy-2'-deoxyguanosine (8-OHdG) using 8-OHdG-aptamer (Apt) as the recognition element. The Apt was adsorbed onto the surface of gold nanoparticles (AuNPs), which prevents them from aggregating under high-salt conditions. Upon addition of 8-OHdG, the conformation of the Apt changes to form a G-quadruplex structure, which leads to the aggregation of the AuNPs along with the increase of the resonance light scattering intensity. The mechanism of 8-OHdG that induces Apt to form G-quadruplexes structure was studied by circular dichroism. The response signals linearly correlated with the concentration of 8-OHdG ranging from 90.8pM to 14.1nM with a detection limit of 27.3pM, which is much lower than that obtained by other methods. This method does not need any label steps and sophisticated equipment. The application for detection of 8-OHdG in real samples further demonstrated its reliability. This strategy would be helpful for developing a universal analytical method by simply replacing corresponding aptamers for various target molecules.

摘要

我们开发了一种基于 8-羟基-2'-脱氧鸟苷(8-OHdG)适配体(Apt)作为识别元件的高灵敏度无标记检测方法。Apt 被吸附到金纳米粒子(AuNPs)的表面上,这使得它们在高盐条件下不会聚集。当加入 8-OHdG 时,Apt 的构象发生变化,形成 G-四链体结构,导致 AuNPs 聚集并伴随着共振光散射强度的增加。通过圆二色性研究了 8-OHdG 诱导 Apt 形成 G-四链体结构的机制。响应信号与 8-OHdG 的浓度呈线性相关,浓度范围为 90.8pM 至 14.1nM,检测限为 27.3pM,比其他方法低得多。该方法不需要任何标记步骤和复杂的设备。该方法在实际样品中检测 8-OHdG 的应用进一步证明了其可靠性。通过简单地替换相应的适配体,这种策略将有助于开发通用的分析方法,用于各种目标分子。

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