• 文献标题:   Label-Free DNA Biosensor Using Modified Reduced Graphene Oxide Platform as a DNA Methylation Assay
  • 文献类型:   Article
  • 作  者:   SEDLACKOVA E, BYTESNIKOVA Z, BIRGUSOVA E, SVEC P, ASHRAFI AM, ESTRELA P, RICHTERA L
  • 作者关键词:   nanomaterial, electrochemical biosensor, dna methylation, rgo, biomedical application
  • 出版物名称:   MATERIALS
  • ISSN:  
  • 通讯作者地址:   Mendel Univ Brno
  • 被引频次:   0
  • DOI:   10.3390/ma13214936
  • 出版年:   2020

▎ 摘  要

This work reports the use of modified reduced graphene oxide (rGO) as a platform for a label-free DNA-based electrochemical biosensor as a possible diagnostic tool for a DNA methylation assay. The biosensor sensitivity was enhanced by variously modified rGO. The rGO decorated with three nanoparticles (NPs)-gold (AuNPs), silver (AgNPs), and copper (CuNPs)-was implemented to increase the electrode surface area. Subsequently, the thiolated DNA probe (single-stranded DNA, 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 a HpaII endonuclease specific site sequence of CpG (5 '-CCGG-3 ') islands. For monitoring the MTase activity, differential pulse voltammetry (DPV) was used, whereas the best results were obtained by rGO-AuNPs. This assay is rapid, cost-effective, sensitive, selective, highly specific, and displays a low limit of detection (LOD) of 0.06 U center dot mL(-1). Lastly, this study was enriched with the real serum sample, where a 0.19 U center dot 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.