An electrochemical genosensor for differentiation of fully methylated from fully unmethylated states of BMP3 gene.

Autor: Hadian-Ghazvini S; Laboratory of Bioanalysis, Institute of Biochemistry and Biophysics, University of Tehran, Tehran 1417614335, Iran., Dashtestani F; Laboratory of Bioanalysis, Institute of Biochemistry and Biophysics, University of Tehran, Tehran 1417614335, Iran., Hakimian F; Laboratory of Bioanalysis, Institute of Biochemistry and Biophysics, University of Tehran, Tehran 1417614335, Iran., Ghourchian H; Laboratory of Bioanalysis, Institute of Biochemistry and Biophysics, University of Tehran, Tehran 1417614335, Iran. Electronic address: ghourchian@ut.ac.ir.
Jazyk: angličtina
Zdroj: Bioelectrochemistry (Amsterdam, Netherlands) [Bioelectrochemistry] 2021 Dec; Vol. 142, pp. 107924. Date of Electronic Publication: 2021 Aug 08.
DOI: 10.1016/j.bioelechem.2021.107924
Abstrakt: The methylation state of a part of the BMP3 gene was detected by our genosensor. This epigenetic biomarker is involved in the biomarker panel of the sDNA test, which is an FDA approved test for colorectal cancer screening. In the present genosensor, polyethyleneimine-stabilized silver nanoparticles (PEI-AgNPs) were used as a non-specific nanolabel for signal generation/amplification and lowering the limit of detection. After immobilization of capture probes and mercaptoethanol molecules on the gold electrode, a thermally treated mixture of the BMP3 targets and reporter probes was introduced to the electrode. Because of the specificity of the reporter probes for fully methylated targets, complete sandwich-like complexes are formed only with them. Therefore, such full-length double-stranded hybrids compared to fully unmethylated targets have more negative charges and can more attract positively charged PEI-AgNPs. For discrimination between methylated and unmethylated targets, electroimpedance spectroscopy and cyclic voltammetry were used for electrode modification monitoring and signal measurement. The sharp and narrow anodic peaks of cyclic voltammograms, which resulted from silver oxidation, were utilized for calibration plot analysis. The genosensor showed a linear response for the target concentration range from 1fM to 100 nM, while the detection limit for methylated and unmethylated target discrimination was 1 fM.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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