Graphene sheets, polyaniline and AuNPs based DNA sensor for electrochemical determination of BCR/ABL fusion gene with functional hairpin probe
Graphene sheets, polyaniline and AuNPs based DNA sensor for electrochemical determination of BCR/ABL fusion gene with functional hairpin probe
复制标题
基于石墨烯片、聚苯胺和 AuNPs 的 DNA 传感器，用于使用功能性发夹探针电化学测定 BCR/ABL 融合基因
DOI：
10.1016/j.bios.2013.07.049
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发表时间：
2014-01-15
影响因子：
12.6
通讯作者：
Feng, Wenli
中科院分区：
文献类型：
--
作者：
Wang, Li;Hua, Erhui;Feng, Wenli
研制了一种灵敏、选择性好的电化学DNA传感器，用于检测慢性粒细胞白血病（CML）的BCR/ABL融合基因。首先用壳聚糖（CS）溶液制备石墨烯片（GS）悬浮液，然后将其组装到玻碳电极（GCE）上，再通过苯胺的电聚合形成聚苯胺膜，最后在修饰的玻碳电极上电沉积Au纳米粒子（AuNPs），以修饰捕获探针。捕获探针采用发夹结构，并用5 '-SH和3'-生物素双重标记。在与靶DNA杂交后，发夹结构被迫打开，并且3 '-生物素被迫远离电极表面。结果表明，链霉亲和素-碱性磷酸酶（SA-AP）通过生物素-亲和素系统与捕获探针共价结合。然后在催化电惰性的1-磷酸萘酯（1-NP）水解为1-萘酚后产生还原电流，并通过差分脉冲伏安法（DPV）进行监测。在最佳条件下，电流信号与目标DNA浓度（10 ~ 1000 pM）呈线性关系，检测限低至2.11 pM（S/N=3），具有良好的区分能力，该方法具有良好的稳定性和重现性。用于真实的样品中BCR/ABL融合基因的检测，结果满意。(C)2013爱思唯尔有限公司版权所有。
A sensitive and selective electrochemical DNA sensor was developed for the detection of BCR/ABL fusion gene in chronic myelogenous leukemia (CML). Firstly, graphene sheets (GS) suspension was prepared with the aid of chitosan (CS) solution and then fabricated onto the glassy carbon electrode (GCE), followed by the electro-polymerization of aniline to form the PANI layer, then, Au nanoparticles (AuNPs) were electro-deposited onto the modified GCE to immobilize the capture probes. The capture probe employed a hairpin structure and dually labeled with a 5'-SH and a 3'-biotin. After hybridization with the target DNA, hairpin structure was compelled to open and 3'-biotin was forced to stay away from the electrode surface. As a result, streptavidin-alkaline phosphatase (SA-AP) was covalently binded to the capture probe via biotin-avidin system. Reduction currents were then generated after catalyzing the hydrolysis of the electroinactive 1-naphthyl phosphate (1-NP) to 1-naphthol and monitored by differential pulse voltammetry (DPV). Under optimum conditions, the amperometric signals increased linearly with the target DNA concentrations (10 pM to 1000 pM), and the DNA sensor exhibited a detection limit as low as 2.11 pM (S/N=3) with an excellent differentiation ability, and the proposed method showed acceptable stability and reproducibility. It has been applied for assay of BCR/ABL fusion gene from real samples with satisfactory results. (C) 2013 Elsevier B.V. All rights reserved.