Electrochemical genosensor for the direct detection of tailed PCR amplicons incorporating ferrocene labelled dATP
Author
Magriñá, Ivan
Toldrà, Anna
Ortiz, Mayreli
Simonova, Anna
Katakis, Ioanis
Hocek, Michal
O'Sullivan, Ciara K.
Publication date
2019-03-30ISSN
0956-5663
Abstract
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.
Document Type
Article
Document version
Accepted version
Language
English
Subject (CDU)
63 - Agriculture and related sciences and techniques
Pages
21
Publisher
Elsevier
Is part of
Biosensors and Bioelectronics
Citation
Magriñá, Ivan, Anna Toldrà, Mònica Campàs, Mayreli Ortiz, Anna Simonova, Ioanis Katakis, Michal Hocek, and Ciara K. O'Sullivan. 2019. "Electrochemical Genosensor For The Direct Detection Of Tailed PCR Amplicons Incorporating Ferrocene Labelled Datp". Biosensors And Bioelectronics 134: 76-82. Elsevier BV. doi:10.1016/j.bios.2019.03.060.
Grant agreement number
MINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/BIO2014-56024-C2-2-R/ES/MICROSISTEMAS PARA LA DETECCION RAPIDA, FIABLE Y RENTABLE DE MICROALGAS TOXICAS IN SITU Y A TIEMPO REAL/SEASENSING
MINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/BIO2017-87946-C2-2-R/ES/DISPOSITIVOS BIOANALITICOS PARA LA DETECCION RAPIDA Y RENTABLE DE GAMBIERDISCUS Y CIGUATOXINAS/CIGUASENSING
Program
Aigües Marines i Continentals
This item appears in the following Collection(s)
- ARTICLES CIENTÍFICS [2510]
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Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-nd/4.0/