Autor segons l'article: Jauset-Rubio, Miriam; Ortiz, Mayreli; O'Sullivan, Ciara K
Departament: Enginyeria Química
Autor/s de la URV: Jauset Rubio, Miriam / O'SULLIVAN, CIARA KATHLEEN / Ortíz Rodríguez, Mayreli
Paraules clau: Triphosphate Solid-phase Solid phasis Single nucleotide polymorphisms Single nucleotide polymorphism sites Recombinases Recombinase polymerase amplification Proteins Primer extension Polymorphism Optical detection Nucleotides Nucleic-acid amplification Mycobacterium-tuberculosis Mediated isothermal amplification Hypertrophic cardiomyopathy Gold nanoparticles Enzymatic-synthesis Electrochemical detection Dna Combination Blood samples Blood Biosensor
Resum: Isothermal recombinase polymerase amplification-based solid-phase primer extension is used for the optical detection of a hypertrophic cardiomyopathy associated single nucleotide polymorphism (SNP) in a fingerprick blood sample. The assay exploits four thiolated primers which have the same sequences with the exception of the 3'-terminal base. Target DNA containing the SNP site hybridizes to all four of the immobilized probes, with primer extension only taking place from the primer containing the terminal base that is complementary to the SNP under interrogation. Biotinylated deoxynucleotide triphosphates are used in the primer extension, allowing postextension addition of streptavidin-poly-horseradish peroxidase to bind to the incorporated biotinylated dNTPs. The signal generated following substrate addition can then be measured optically. The percentage of biotinylated dNTPs and the duration of primer extension is optimized and the system applied to the identification of a SNP in a fingerprick blood sample. A methodology of thermal lysis using a 1 in 5 dilution of the fingerprick blood sample prior to application of 95 degrees C for 30 s is used to extract genomic DNA, which is directly used as a template for solid-phase primer extension on microtiter plates, followed by optical detection. The SNP in the fingerprick sample was identified and its identity corroborated using ion torrent next generation sequencing. Ongoing work is focused on extension to the multiplexed detection of SNPs in fingerprick and other biological samples.
Àrees temàtiques: Química Medicina ii Medicina i Materiais Interdisciplinar Geociências General medicine Farmacia Ensino Engenharias iv Engenharias iii Engenharias ii Enfermagem Ciências biológicas iii Ciências biológicas ii Ciências biológicas i Ciências ambientais Ciências agrárias i Ciência de alimentos Ciência da computação Chemistry, analytical Biotecnología Biodiversidade Astronomia / física Analytical chemistry
Accès a la llicència d'ús: https://creativecommons.org/licenses/by/3.0/es/
Adreça de correu electrònic de l'autor: mayreli.ortiz@urv.cat miriam.jauset@urv.cat mayreli.ortiz@urv.cat
Identificador de l'autor: 0000-0002-9423-0055 0000-0002-9943-6132 0000-0002-9423-0055
Data d'alta del registre: 2024-10-26
Versió de l'article dipositat: info:eu-repo/semantics/publishedVersion
Enllaç font original: https://pubs.acs.org/doi/10.1021/acs.analchem.1c03419
URL Document de llicència: https://repositori.urv.cat/ca/proteccio-de-dades/
Referència a l'article segons font original: Analytical Chemistry. 93 (44): 14578-14585
Referència de l'ítem segons les normes APA: Jauset-Rubio, Miriam; Ortiz, Mayreli; O'Sullivan, Ciara K (2021). Solid-Phase Primer Elongation Using Biotinylated dNTPs for the Detection of a Single Nucleotide Polymorphism from a Fingerprick Blood Sample. Analytical Chemistry, 93(44), 14578-14585. DOI: 10.1021/acs.analchem.1c03419
DOI de l'article: 10.1021/acs.analchem.1c03419
Entitat: Universitat Rovira i Virgili
Any de publicació de la revista: 2021
Tipus de publicació: Journal Publications