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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F20%3A00524530" target="_blank" >RIV/68081707:_____/20:00524530 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:14740/20:00118310

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S157266572030134X?via%3Dihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S157266572030134X?via%3Dihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.jelechem.2020.113951" target="_blank" >10.1016/j.jelechem.2020.113951</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10405 - Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis)

Projekt

<a href="/cs/project/GAP206%2F11%2F1638" target="_blank" >GAP206/11/1638: Nové elektrochemické senzory a detekční techniky pro analýzu struktury a interakcí nukleových kyselin</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Electroanalytical Chemistry

ISSN

1572-6657

e-ISSN

—

Svazek periodika

862

Číslo periodika v rámci svazku

APR 1 2020

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

7

Strana od-do

113951

Kód UT WoS článku

000525903900004

EID výsledku v databázi Scopus

2-s2.0-85080093599