Label-free DNA biosensor using modified reduced graphene oxide platform as a DNA methylation assay

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dc.contributor.authorDostálová, Eliškacs
dc.contributor.authorBytešníková, Zuzanacs
dc.contributor.authorBirgusová, Eliškacs
dc.contributor.authorŠvec, Pavelcs
dc.contributor.authorAshrafi, Amirmansoorcs
dc.contributor.authorEstrela, Pedrocs
dc.contributor.authorRichtera, Lukášcs
dc.coverage.issue21cs
dc.coverage.volume13cs
dc.date.issued2020-11-30cs
dc.description.abstractThis 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.en
dc.description.abstractThis 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.en
dc.formattextcs
dc.format.extent1-12cs
dc.format.mimetypeapplication/pdfcs
dc.identifier.citationMaterials. 2020, vol. 13, issue 21, p. 1-12.en
dc.identifier.doi10.3390/ma13214936cs
dc.identifier.issn1996-1944cs
dc.identifier.orcid0000-0003-1367-9414cs
dc.identifier.orcid0000-0002-8288-3999cs
dc.identifier.other168307cs
dc.identifier.researcheridN-9991-2014cs
dc.identifier.scopus12040049600cs
dc.identifier.urihttp://hdl.handle.net/11012/195920
dc.language.isoencs
dc.publisherMDPIcs
dc.relation.ispartofMaterialscs
dc.relation.urihttps://www.mdpi.com/1996-1944/13/21/4936cs
dc.rightsCreative Commons Attribution 4.0 Internationalcs
dc.rights.accessopenAccesscs
dc.rights.sherpahttp://www.sherpa.ac.uk/romeo/issn/1996-1944/cs
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/cs
dc.subjectnanomaterialsen
dc.subjectelectrochemical biosensoren
dc.subjectDNA methylationen
dc.subjectrGOen
dc.subjectbiomedical applicationsen
dc.subjectnanomaterials
dc.subjectelectrochemical biosensor
dc.subjectDNA methylation
dc.subjectrGO
dc.subjectbiomedical applications
dc.titleLabel-free DNA biosensor using modified reduced graphene oxide platform as a DNA methylation assayen
dc.title.alternativeLabel-free DNA biosensor using modified reduced graphene oxide platform as a DNA methylation assayen
dc.type.driverarticleen
dc.type.statusPeer-revieweden
dc.type.versionpublishedVersionen
sync.item.dbidVAV-168307en
sync.item.dbtypeVAVen
sync.item.insts2025.10.14 15:16:42en
sync.item.modts2025.10.14 09:53:05en
thesis.grantorVysoké učení technické v Brně. Středoevropský technologický institut VUT. Chytré nanonástrojecs
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