An electrochemical aptasensor based on hybridization chain reaction with enzyme-signal amplification for interferon-gamma detection
An electrochemical aptasensor based on hybridization chain reaction with enzyme-signal amplification for interferon-gamma detection
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基于杂交链式反应和酶信号放大的电化学适体传感器用于干扰素γ检测
DOI：
10.1016/j.bios.2012.04.013
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发表时间：
2012-06-01
影响因子：
12.6
通讯作者：
Yu, Ruqin
中科院分区：
文献类型：
--
作者：
Zhao, Jingjin;Chen, Chunfei;Yu, Ruqin
构建了一种基于杂交链式反应（HCR）和酶信号放大的新型电化学适体传感器，用于检测干扰素-γ（IFN-γ）。在该适体传感器中，含有IFN-γ适体序列的识别探针最初与IFN-γ结合。未结合的识别探针被捕获在电极上作为引发剂来触发HCR。两个DNA发夹bio-H1和bio-H2被识别探针打开，并一一结合在电极上。生物素用作发夹中的示踪剂，链霉亲和素-碱性磷酸酶（SA-ALP）用作报告分子。然后，SA-ALP 将其电惰性底物 1-萘基磷酸盐转化为电活性衍生物 1-萘酚，通过微分脉冲伏安法 (DPV) 产生放大的电化学信号。通过测量1-萘基磷酸酶促去磷酸化产生的1-萘酚的量，以伏安法测定固定化酶的活性。观察到的电化学信号与IFN-γ的浓度成反比。所提出的方法对浓度范围为 0.5-300 nM 的 IFN-γ 显示出高灵敏度，检测限为 0.3 nM。该传感系统还为细胞培养基中 IFN-γ 的检测提供了令人满意的结果。 (C) 2012 Elsevier B.V. 保留所有权利。
A novel electrochemical aptasensor based on hybridization chain reaction (HCR) with enzyme-signal amplification was constructed for the detection of interferon-gamma (IFN-gamma). In this aptasensor, the recognition probes which contained the sequence of IFN-gamma aptamer were initially binded to IFN-gamma. and the unbound recognition probes were captured on the electrode as an initiator to trigger the HCR. The two DNA hairpins bio-H1 and bio-H2 were opened by the recognition probe, and bound one by one on the electrode. The biotin was used as a tracer in the hairpins and streptavidin-alkaline phosphatase (SA-ALP) as a reporter molecule. Then, SA-ALP converted its electro-inactive substrate 1-naphthyl phosphate into an electroactive derivative 1-naphthol generating amplified electrochemical signal by differential pulse voltammetry (DPV). The activity of the immobilized enzyme was voltammetrically determined by measuring the amount of 1-naphthol generated for enzymatic dephosphorylation of 1-naphthyl phosphate. The electrochemical signal observed was inversely related to the concentration of IFN-gamma. The proposed approach showed a high sensitivity for IFN-gamma in a concentration range of 0.5-300 nM with a detection limit of 0.3 nM. The sensing system also provided satisfactory results for the detection of IFN-gamma in the cell media. (C) 2012 Elsevier B.V. All rights reserved.