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Fast enzyme-linked electrochemical sensing of DNA hybridization at pencil graphite electrodes. Application to detect gene deletion in a human cell culture
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SYSNO ASEP 0524530 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Fast enzyme-linked electrochemical sensing of DNA hybridization at pencil graphite electrodes. Application to detect gene deletion in a human cell culture Author(s) Špaček, Jan (BFU-R) ORCID
Eksin, E. (TR)
Havran, Luděk (BFU-R) RID, ORCID
Erdem, A. (TR)
Fojta, Miroslav (BFU-R) RID, ORCIDNumber of authors 5 Article number 113951 Source Title Journal of Electroanalytical Chemistry. - : Elsevier - ISSN 1572-6657
Roč. 862, APR 1 2020 (2020)Number of pages 7 s. Publication form Online - E Language eng - English Country CH - Switzerland Keywords single nucleotide polymorphisms ; sensitive detection ; genomagnetic assay ; pcr products ; biosensor Subject RIV CG - Electrochemistry OECD category Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis) R&D Projects GAP206/11/1638 GA ČR - Czech Science Foundation (CSF) Method of publishing Limited access Institutional support BFU-R - RVO:68081707 UT WOS 000525903900004 EID SCOPUS 85080093599 DOI 10.1016/j.jelechem.2020.113951 Annotation In this paper we present a rapid electrochemical enzyme-linked DNA hybridization assay using disposable pencil graphite electrodes (PeGE) to detect target DNA (tDNA) sequences in DNA fragments amplified by polymerase chain reaction. The procedure consists of several short (1-2 min) incubation steps, including adsorption of the tDNA at unpretreated PeGE from denaturing medium, surface blocking with milk proteins, hybridization with a biotinylated oligonucleotide probe and binding of streptavidin-alkaline phosphatase conjugate to the biotin tags. Then the PeGE is transferred into background electrolyte solution containing 1-naphthyl phosphate, which is enzymatically dephosphorylated to give electrochemically oxidizable indicator 1-naphthol. The assay, which can be performed within 7-8 min, offers a perfect discrimination between specific and nonspecific DNA amplicons and easy detection of about similar to 40 femtomoles of tDNA in large excesses of non-complementary DNA. An application on the detection of p53 gene deletion in a human cell culture, featuring a real biologicalmodel, is presented. The designed setup has a potential to be applied as one of simple, fast, robust ultralow cost do-it-yourself instruments recently introduced as diagnostic tools for third world countries. (c) 2020 Elsevier B.V. All rights reserved. Workplace Institute of Biophysics Contact Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Year of Publishing 2021 Electronic address https://www.sciencedirect.com/science/article/pii/S157266572030134X?via%3Dihub
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