Electrochemical genosensor for the direct detection of tailed PCR amplicons incorporating ferrocene labelled dATP

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F19%3A00504622" target="_blank" >RIV/61388963:_____/19:00504622 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11310/19:10394787

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S0956566319302751?via%3Dihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0956566319302751?via%3Dihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.bios.2019.03.060" target="_blank" >10.1016/j.bios.2019.03.060</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Electrochemical genosensor for the direct detection of tailed PCR amplicons incorporating ferrocene labelled dATP

Popis výsledku v původním jazyce

An electrochemical genosensor for the detection and quantification of Karlodinium armiger is presented. The genosensor exploits tailed primers and ferrocene labelled dATP analogue to produce PCR products that can be directly hybridised on a gold electrode array and quantitatively measured using square wave voltammetry. Tailed primers consist of a sequence specific for the target, followed by a carbon spacer and a sequence specifically designed not to bind to genomic DNA, resulting in a duplex flanked by single stranded binding primers. The incorporation of the 7-(ferrocenylethynyl)-7-deaza-2'-deoxyadenosine triphosphate was optimised in terms of a compromise between maximum PCR efficiency and the limit of detection and sensitivity attainable using electrochemical detection via hybridisation of the tailed, ferrocene labelled PCR product. A limit of detection of 277aM with a linear range from 315aM to 10 fM starting DNA concentration and a sensitivity of 122 nA decade(-1) was achieved. The system was successfully applied to the detection of genomic DNA in real seawater samples.

Název v anglickém jazyce

Electrochemical genosensor for the direct detection of tailed PCR amplicons incorporating ferrocene labelled dATP

Popis výsledku anglicky

An electrochemical genosensor for the detection and quantification of Karlodinium armiger is presented. The genosensor exploits tailed primers and ferrocene labelled dATP analogue to produce PCR products that can be directly hybridised on a gold electrode array and quantitatively measured using square wave voltammetry. Tailed primers consist of a sequence specific for the target, followed by a carbon spacer and a sequence specifically designed not to bind to genomic DNA, resulting in a duplex flanked by single stranded binding primers. The incorporation of the 7-(ferrocenylethynyl)-7-deaza-2'-deoxyadenosine triphosphate was optimised in terms of a compromise between maximum PCR efficiency and the limit of detection and sensitivity attainable using electrochemical detection via hybridisation of the tailed, ferrocene labelled PCR product. A limit of detection of 277aM with a linear range from 315aM to 10 fM starting DNA concentration and a sensitivity of 122 nA decade(-1) was achieved. The system was successfully applied to the detection of genomic DNA in real seawater samples.

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/EF16_019%2F0000729" target="_blank" >EF16_019/0000729: Chemická biologie pro vývoj nových terapií</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

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

Název periodika

Biosensors and Bioelectronics

ISSN

0956-5663

e-ISSN

—

Svazek periodika

134

Číslo periodika v rámci svazku

Jun 1

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

7

Strana od-do

76-82

Kód UT WoS článku

000466248400011

EID výsledku v databázi Scopus

2-s2.0-85063760535