Author, as appears in the article.: Ortiz, Mayreli; Jauset-Rubio, Miriam; Kodr, David; Simonova, Anna; Hocek, Michal; O'Sullivan, Ciara K
Department: Enginyeria Química
URV's Author/s: Jauset Rubio, Miriam / O'SULLIVAN, CIARA KATHLEEN / Ortíz Rodríguez, Mayreli
Keywords: Solid-phase Solid phasis Solid phase recombinase polymerase amplification Snp Recombinases Primer extension Organometallics Oligonucleotides Iron compounds Fingerprick Ferrocenes Ferrocene-dntps Ferrocene-dntp Ferrocene labelled Electrodes Electrochemical detection Dna Chemical detection Blood Biosensors solid phase recombinase polymerase snp pcr mutation detection multiplex gold genomic dna ferrocene-dntps electrochemical detection chip chain-reaction biosensors assay array amplification
Abstract: Hypertrophic cardiomyopathies (HCM) are the principal cause of sudden cardiac death in young athletes and it is estimated that 1 in 500 people have HCM. The aim of this work was to develop an electrochemical platform for the detection of HCM-associated SNP in the Myosin Heavy Chain 7 (MYH7) gene, in fingerprick blood samples. The platform exploits isothermal solid-phase primer elongation using recombinase polymerase amplification with either individual or a combination of four ferrocene-labelled nucleoside triphosphates. Four thiolated reverse primers containing a variable base at their 3’ end were immobilised on individual gold electrodes of an array. Following hybridisation with target DNA, solid phase recombinase polymerase amplification was carried out and primer elongation incorporating the ferrocene labelled oligonucleotides was only detected at one of the electrodes, thus facilitating identification of the SNP under interrogation. The assay was applied to the direct detection of the SNP in fingerprick blood samples from eight different individuals, with the results obtained corroborating with next generation sequencing. The ability to be able to robustly identify the SNP using a 10 ?L fingerprick sample, demonstrates that SNP discrimination is achieved using low femtomolar (ca. 8 × 105 copies DNA) levels of DNA. © 2021 The Authors
Thematic Areas: Química Nanoscience and nanotechnology Nanoscience & nanotechnology Medicine (miscellaneous) Medicina ii Medicina i Materiais Interdisciplinar Farmacia Engenharias iv Engenharias iii Engenharias ii Electrochemistry Economia Ciências biológicas iii Ciências biológicas ii Ciências biológicas i Ciências ambientais Ciências agrárias i Chemistry, analytical Biotecnología Biotechnology & applied microbiology Biotechnology Biophysics Biomedical engineering Biodiversidade Astronomia / física
licence for use: https://creativecommons.org/licenses/by/3.0/es/
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://www.sciencedirect.com/science/article/pii/S0956566321008629?via%3Dihub
Licence document URL: https://repositori.urv.cat/ca/proteccio-de-dades/
Papper original source: Biosensors & Bioelectronics. 198 113825-
APA: Ortiz, Mayreli; Jauset-Rubio, Miriam; Kodr, David; Simonova, Anna; Hocek, Michal; O'Sullivan, Ciara K (2022). Solid-phase recombinase polymerase amplification using ferrocene-labelled dNTPs for electrochemical detection of single nucleotide polymorphisms. Biosensors & Bioelectronics, 198(), 113825-. DOI: 10.1016/j.bios.2021.113825
Article's DOI: 10.1016/j.bios.2021.113825
Entity: Universitat Rovira i Virgili
Journal publication year: 2022
Publication Type: Journal Publications
Repositori URV