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. 2022 Oct 17:2022:6575140.
doi: 10.1155/2022/6575140. eCollection 2022.

Development of a Nanostructured Electrochemical Genosensor for the Detection of the K-ras Gene

Luis Fernando Garcia-Melo  1   2 , Miguel Morales-Rodríguez  1 , Eduardo Madrigal-Bujaidar  3 , Eduardo O Madrigal-Santillán  4 , José Antonio Morales-González  4 , Rosa Natali Pineda Cruces  2 , Jorge Alfredo Campoy Ramírez  2 , Pablo Damian-Matsumura  5 , Alexandro Tellez-Plancarte  3 , Nikola Batina  2 , Isela Álvarez-González  3
Affiliations

Development of a Nanostructured Electrochemical Genosensor for the Detection of the K-ras Gene

Luis Fernando Garcia-Melo et al. J Anal Methods Chem. .

Abstract

In the scientific literature, it has been documented that electrochemical genosensors are novel analytical tools with proven clinical diagnostic potential for the identification of carcinogenic processes due to genetic and epigenetic alterations, as well as infectious diseases due to viruses or bacteria. In the present work, we describe the construction of an electrochemical genosensor for the identification of the k12p.1 mutation; it was based on use of Screen-Printed Gold Electrode (SPGE), Cyclic Voltammetry (CV), and Atomic Force Microscopy (AFM), for the monitoring the electron transfer trough the functionalized nanostructured surface and corresponding morphological changes. The sensitivity of the genosensor showed a linear response for the identification of the k12p.1 mutation of the K-ras gene in the concentration range of 10 fM to 1 μM with a detection limit of 7.96 fM in the presence of doxorubicin (Dox) as DNA intercalating agent and indicator of the hybridization reaction. Thus, the electrochemical genosensor developed could be useful for the identification of diseases related with the K-ras oncogene.

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Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
Cyclic voltammograms current of the 20 mM [Fe(CN)6]−3/−4 redox probe/0.01 M PBS (pH 7.4) and AFM image of 3D morphology of (for both cases) registered on unmodified Au electrode (a), modified Au/MUCS electrode (b) modified Au/MERS electrode and modified Au/MCHS electrode, with area of 1 μm × 1 μm and Z scale 0–250 nm.
Figure 2
Figure 2
Cyclic voltammograms current of the 20 mM [Fe(CN)6]−3/−4 redox probe/0.01 M PBS (pH 7.4) registered on modified Au/MUCS (a), 10−6 M Au/MUCS/dsDNA electrode (b), and Au/MUCS/dsDNA/Dox electrode (c). AFM image of 3D morphology of registered on Au/MERS/dsDNA electrode (b) and modified Au/MCHS/dsDNA/Dox electrode (c), with area of 1 μm × 1 μm and Z scale 0–250 nm.
Figure 3
Figure 3
(a) Anodic peak current for [Fe(CN)6]3−/4− redox probe recorded at Au/MUCS/dsDNA and Au/MUCS/dsDNA/dox electrodes, in different concentration of target DNA, respectively. (b) Linear relationship between the relative current change of the anodic peak [Fe(CN)6]3−/4− redox probe and the logarithm of the target DNA concentration at Au/MUCS/dsDNA and Au/MUCS/dsDNA/Dox electrodes, respectively.
Figure 4
Figure 4
(a) The relative current change of the anodic peak of [Fe(CN)6]−3/−4 redox probe versus hybridization temperature after the dox treatment, at the room temperature (20°C) (Au/MUCS/dsDNA/dox). (b) Cyclic voltammograms of [Fe(CN)6]3−/4− redox probe recorded at Au/MUCS electrode (a), Au/MUCS/sNC electrode (b), and Au/MUCS/dsDNA electrode (c). (c) Cyclic voltammograms of [Fe(CN)6]3−/4− redox probe recorded at Au/MUCS electrode (a) Au/MUCS/sNC/dox electrode (b), and Au/MUCS/dsDNA/dox electrode. The dox solution was added in excess 10−5 (M) all electrochemical measures were made by means of cyclic voltammetry (CV) at a scan rate of 50 mV/s, within a potential range from −400 mV to +500 mV.
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