Author, as appears in the article.: Magrina, Ivan; Jauset-Rubio, Miriam; Ortiz, Mayreli; Tomaso, Herbert; Simonova, Anna; Hocek, Michal; O'Sullivan, Ciara K
Department: Enginyeria Química
URV's Author/s: Jauset Rubio, Miriam / Magriñá Lobato, Ivan / O'SULLIVAN, CIARA KATHLEEN / Ortíz Rodríguez, Mayreli
Keywords: Triphosphates Redox label Real Number Nucleotides Identification Gene Francisella-tularensis Assays Amplification
Abstract: We report the duplex amplification of two plasmid DNA markers involved in the virulence of Bacillus anthracis, CAP and PAG, and the direct electrochemical detection of these amplicons. The method consists of the simultaneous amplification of the two targets in a single-pot reaction via polymerase chain reaction (PCR) using tailed primers and ferrocene-labeled dATP. Following amplification, the PCR products hybridize to probes immobilized on electrodes in a microfabricated electrode array chip. The incorporated ferrocene labeled dATP is then detected using square wave voltammetry. We evaluated the effect of electrolyte cations, anions, and concentration to condense, bend, and shrink double-stranded DNA and their effect on the intensity of the ferrocene signal. We obtained detection limits 40.8 and 3.4 fM for CAP and PAG targets, respectively. We successfully developed a method to detect the presence of both targets in genomic DNA extracted from real samples.
Thematic Areas: Química Interdisciplinar General chemistry General chemical engineering Engenharias ii Ciências agrárias i Chemistry, multidisciplinary Chemistry (miscellaneous) Chemistry (all) Chemical engineering (miscellaneous) Chemical engineering (all)
licence for use: https://creativecommons.org/licenses/by/3.0/es/
ISSN: 24701343
Author's mail: mayreli.ortiz@urv.cat miriam.jauset@urv.cat mayreli.ortiz@urv.cat
Author identifier: 0000-0002-9423-0055 0000-0002-9943-6132 0000-0002-9423-0055
Record's date: 2024-10-26
Papper version: info:eu-repo/semantics/publishedVersion
Link to the original source: https://pubs.acs.org/doi/abs/10.1021/acsomega.9b02890
Licence document URL: https://repositori.urv.cat/ca/proteccio-de-dades/
Papper original source: Acs Omega. 4 (26): 21900-21908
APA: Magrina, Ivan; Jauset-Rubio, Miriam; Ortiz, Mayreli; Tomaso, Herbert; Simonova, Anna; Hocek, Michal; O'Sullivan, Ciara K (2019). Duplex Electrochemical DNA Sensor to Detect Bacillus anthracis CAP and PAG DNA Targets Based on the Incorporation of Tailed Primers and Ferrocene-Labeled dATP. Acs Omega, 4(26), 21900-21908. DOI: 10.1021/acsomega.9b02890
Article's DOI: 10.1021/acsomega.9b02890
Entity: Universitat Rovira i Virgili
Journal publication year: 2019
Publication Type: Journal Publications
Repositori URV