Fast enzyme-linked electrochemical sensing of DNA hybridization
at pencil graphite electrodes. Application to detect gene
deletion in a human cell culture

J 2020

Fast enzyme-linked electrochemical sensing of DNA hybridization at pencil graphite electrodes. Application to detect gene deletion in a human cell culture

ŠPAČEK, Jan, E,. EKSIN, L. HAVRAN, A. ERDEM, Miroslav FOJTA et. al.

Základní údaje

Originální název

Fast enzyme-linked electrochemical sensing of DNA hybridization at pencil graphite electrodes. Application to detect gene deletion in a human cell culture

Autoři

ŠPAČEK, Jan (203 Česká republika, domácí), E,. EKSIN, L. HAVRAN, A. ERDEM a Miroslav FOJTA (203 Česká republika, garant, domácí)

Vydání

Journal of Electroanalytical Chemistry, Lausanne, Elsevier, 2020, 1572-6657

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10405 Electrochemistry

Stát vydavatele

Švýcarsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.464

Kód RIV

RIV/00216224:14740/20:00118310

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1016/j.jelechem.2020.113951

UT WoS

000525903900004

Klíčová slova anglicky

DNA hybridization; Enzyme-linked electrochemical assay; Gene deletion; Pencil graphite electrode; Ultralow cost; Do-it-yourself devices

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 11. 3. 2021 17:46, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

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.

Citovat

ŠPAČEK, Jan, E,. EKSIN, L. HAVRAN, A. ERDEM a Miroslav FOJTA. Fast enzyme-linked electrochemical sensing of DNA hybridization at pencil graphite electrodes. Application to detect gene deletion in a human cell culture. Journal of Electroanalytical Chemistry. Lausanne: Elsevier, 2020, roč. 862, APR, s. 113951-113957. ISSN 1572-6657. Dostupné z: https://dx.doi.org/10.1016/j.jelechem.2020.113951.

@article{1748117,
   author = {Špaček, Jan and Eksin, E,. and Havran, L. and Erdem, A. and Fojta, Miroslav},
   article_location = {Lausanne},
   article_number = {APR},
   doi = {http://dx.doi.org/10.1016/j.jelechem.2020.113951},
   keywords = {DNA hybridization; Enzyme-linked electrochemical assay; Gene deletion; Pencil graphite electrode; Ultralow cost; Do-it-yourself devices},
   language = {eng},
   issn = {1572-6657},
   journal = {Journal of Electroanalytical Chemistry},
   title = {Fast enzyme-linked electrochemical sensing of DNA hybridization at pencil graphite electrodes. Application to detect gene deletion in a human cell culture},
   url = {https://linkinghub.elsevier.com/retrieve/pii/S157266572030134X},
   volume = {862},
   year = {2020}
}

TY  - JOUR
ID  - 1748117
AU  - Špaček, Jan - Eksin, E,. - Havran, L. - Erdem, A. - Fojta, Miroslav
PY  - 2020
TI  - Fast enzyme-linked electrochemical sensing of DNA hybridization at pencil graphite electrodes. Application to detect gene deletion in a human cell culture
JF  - Journal of Electroanalytical Chemistry
VL  - 862
IS  - APR
SP  - 113951
EP  - 113951
PB  - Elsevier
SN  - 15726657
KW  - DNA hybridization
KW  - Enzyme-linked electrochemical assay
KW  - Gene deletion
KW  - Pencil graphite electrode
KW  - Ultralow cost
KW  - Do-it-yourself devices
UR  - https://linkinghub.elsevier.com/retrieve/pii/S157266572030134X
N2  - 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.
ER  -

ŠPAČEK, Jan, E,. EKSIN, L. HAVRAN, A. ERDEM a Miroslav FOJTA. Fast enzyme-linked electrochemical sensing of DNA hybridization at pencil graphite electrodes. Application to detect gene deletion in a human cell culture. \textit{Journal of Electroanalytical Chemistry}. Lausanne: Elsevier, 2020, roč.~862, APR, s.~113951-113957. ISSN~1572-6657. Dostupné z: https://dx.doi.org/10.1016/j.jelechem.2020.113951.

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