Electrochemical genosensor based on colloidal gold nanoparticles for the detection of Factor V Leiden mutation using disposable pencil graphite electrodes

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dc.contributor.authorOzsoz, M
dc.contributor.authorErdem, A
dc.contributor.authorKerman, K
dc.contributor.authorOzkan, D
dc.contributor.authorTugrul, B
dc.contributor.authorTopcuoglu, N
dc.contributor.authorEkren, H
dc.contributor.authorTaylan, M
dc.date.accessioned2019-10-27T19:00:41Z
dc.date.available2019-10-27T19:00:41Z
dc.date.issued2003
dc.departmentEge Üniversitesien_US
dc.description.abstractElectrochemical genosensors for the detection of the Factor V Leiden mutation from polymerase chain reaction (PCR) amplicons using the oxidation signal of colloidal gold (Au) is described. A pencil graphite electrode (PGE) modified with target DNA, when hybridized with complementary probes conjugated to Au nanoparticles, responded with the appearance of a Au oxide wave at similar to+ 1.20 V. Specific probes were immobilized onto the Au nanoparticles in two different modes: (a) Inosine-substituted probes were covalendy attached from their amino groups at the 5' end using N-(3-dimethylamino)propyl)-N'-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (NHS) as a coupling agent onto a carboxylate-terminated L-cysteine self-assembled monolayer (SAM) preformed on the Au nanoparticles, and (b) probes with a hexanethiol group at their 5' phosphate end formed a SAM on Au nanoparticles. The genosensor relies on the hybridization of the probes with their complementary targets, which are covalendy immobilized at the PGE surface. Au-tagged 23-mer capture probes were challenged with the synthetic 23-mer target, 131-base single-stranded DNA or denatured 256-base polymerase chain reaction (PCR) amplicon. The appearance of the Au oxidation signal shortened the assay time and simplified the detection of the Factor V Leiden mutation from PCR amplified real samples. The discrimination between the homozygous and heterozygous mutations was also established by comparing the peak currents of the Au signals. Numerous factors affecting the hybridization and nonspecific binding events were optimized. The detection limit for the PCR amplicons was found to be as low as 0.78 fmol; thus, it is suitable for point-of-care applications.en_US
dc.identifier.doi10.1021/ac026212ren_US
dc.identifier.endpage2187en_US
dc.identifier.issn0003-2700
dc.identifier.issue9en_US
dc.identifier.pmid12720360en_US
dc.identifier.scopusqualityN/Aen_US
dc.identifier.startpage2181en_US
dc.identifier.urihttps://doi.org/10.1021/ac026212r
dc.identifier.urihttps://hdl.handle.net/11454/37810
dc.identifier.volume75en_US
dc.identifier.wosWOS:000182662800045en_US
dc.identifier.wosqualityQ1en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherAmer Chemical Socen_US
dc.relation.ispartofAnalytical Chemistryen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.titleElectrochemical genosensor based on colloidal gold nanoparticles for the detection of Factor V Leiden mutation using disposable pencil graphite electrodesen_US
dc.typeArticleen_US

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