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基于双功能石墨烯衍生物的电化学平台用于在生物样品中检测具有单核苷酸多态性选择性的TP53基因。

Dual functional graphene derivative-based electrochemical platforms for detection of the TP53 gene with single nucleotide polymorphism selectivity in biological samples.

作者信息

Esteban-Fernández de Ávila Berta, Araque Elena, Campuzano Susana, Pedrero María, Dalkiran Berna, Barderas Rodrigo, Villalonga Reynaldo, Kiliç Esma, Pingarrón José M

机构信息

Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid , E-28040 Madrid, Spain.

出版信息

Anal Chem. 2015 Feb 17;87(4):2290-8. doi: 10.1021/ac504032d. Epub 2015 Jan 30.

Abstract

Novel disposable electrochemical DNA sensors were prepared for the detection of a target DNA sequence on the p53 tumor suppressor (TP53) gene. The electrochemical platform consisted of screen-printed carbon electrodes (SPCEs) functionalized with a water-soluble reduced graphene oxide-carboxymethylcellulose (rGO-CMC) hybrid nanomaterial. Two different configurations involving hairpin specific capture probes of different length covalently immobilized through carbodiimide chemistry on the surface of rGO-CMC-modified SPCEs were implemented and compared. Upon hybridization, a streptavidin-peroxidase (Strep-HRP) conjugate was employed as an electrochemical indicator. Hybridization was monitored by recording the amperometric responses measured at -0.10 V (vs an Ag pseudo-reference electrode) upon the addition of 3,3',5,5'-tetramethylbenzidine (TMB) as a redox mediator and H2O2 as an enzyme substrate. The implemented DNA platforms allow single nucleotide polymorphism (SNP) discrimination in cDNAs from human breast cancer cell lines, which makes such platforms excellent as new diagnosis tools in clinical analysis.

摘要

制备了新型一次性电化学DNA传感器,用于检测p53肿瘤抑制基因(TP53)上的目标DNA序列。该电化学平台由用可水溶性还原氧化石墨烯-羧甲基纤维素(rGO-CMC)杂化纳米材料功能化的丝网印刷碳电极(SPCE)组成。实施并比较了两种不同的配置,其中涉及通过碳二亚胺化学方法将不同长度的发夹特异性捕获探针共价固定在rGO-CMC修饰的SPCE表面。杂交后,使用链霉亲和素-过氧化物酶(Strep-HRP)偶联物作为电化学指示剂。通过记录在添加作为氧化还原介质的3,3',5,5'-四甲基联苯胺(TMB)和作为酶底物的H2O2后在-0.10 V(相对于Ag伪参比电极)下测量的安培响应来监测杂交。所实施的DNA平台能够区分来自人乳腺癌细胞系的cDNA中的单核苷酸多态性(SNP),这使得此类平台成为临床分析中出色的新型诊断工具。

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