Label-Free DNA Biosensor using Modified Reduced Graphene Oxide Platform as a DNA Methylation Assay

Eliska Sedlackova, Zuzana Bytesnikova, Eliska Birgusova, Pavel Svec, Amirmansoor Ashrafi, Pedro Estrela, Lukas Richtera

Research output: Contribution to journalArticle

Abstract

This work reports using modified reduced graphene oxide (rGO) as a platform for label-free DNA-based electrochemical biosensor as a possible diagnostic tool for DNA methylation assay. The biosensor sensitivity wasenhanced by variously modified rGO. The rGO decorated with three nanoparticles (NPs)—gold (AuNPs), silver (AgNPs), and copper (CuNPs) were implemented to increase the electrode surface area. Subsequently, the thiolated DNA probe (ssDNA−1) was hybridized with the target DNA sequence (ssDNA-2). After the hybridization, the double-stranded DNA (dsDNA) was methylated by M.SssI methyltransferase (MTase) and then digested via HpaII endonuclease specific site sequence of CpG islands 5’-CCGG-3’. For the monitoring of the MTase activity was used differential pulse voltammetry (DPV), whereas the best results were obtained by rGO-AuNPs. This assay is rapid, cost-effective, sensitive, selective, highly specific, and displays a low detection limit (LOD) of 0.06 U·mL−1. Lastly, this study was enriched with the real serum sample, where a 0.19 U·mL−1 LOD was achieved. Moreover, the developed biosensor offers excellent potential in future applications in clinical diagnostics, as this approach can be used in the design of other biosensors.
Original languageEnglish
Article number4936
Number of pages12
JournalMaterials
Volume13
Issue number21
DOIs
Publication statusPublished - 3 Nov 2020

Keywords

  • Nanomaterials
  • Electrochemical Biosensor
  • DNA Methylation
  • Reduced graphene oxide
  • biomedical applications

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