Dual Functional Graphene Derivative-Based Electrochemical Platforms for Detection of the TP53 Gene with Single Nucleotide Polymorphism Selectivity in Biological Samples

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.

Electrical Property of Graphene and Its Application to Electrochemical Biosensing

“…Esteban-Fernández de Ávila et al designed a disposable electrochemical DNA sensor based on carboxymethyl-cellulose-rGO modified screen-printed carbon electrodes. And HRP was utilized to catalyze the redox mediator, tetramethylbenzidine (TMB), and the substrate (H 2 O 2 ) for the detection of the p53 tumor suppressor gene [73]. Besides, graphene can be also utilized for DNA sequencing.…”

The Translational Potential of Electrochemical DNA-Based Liquid Biopsy

“…Saliva analysis is performed after minimal pretreatment in 10 min with a LOD of 3.9 fM for mRNA (Wei et al, 2009). The use of undiluted saliva requires proper antifouling strategies while admitting high target binding capacity as demonstrated with a hybrid nanomaterial composed of reduced graphene oxide and carboxymethylcellulose (Esteban-Fernandez de Avila et al, 2015). Discrimination of a single central mismatched TP53 sequence was possible in spiked saliva, but at relatively high concentrations.…”

Electrochemical biosensors based on nanomodified screen-printed electrodes: Recent applications in clinical analysis

“…Among these, many are those based on different signal amplification procedures that allow to achieve very low detection limits [72][73][74][75][76][77][78][79][80]. This, however, generally requires the addition of exogenous, label containing secondary probes and, typically multi-component deposition/amplification steps.…”