Autor segons l'article: AL-Madhagi, Sallam; O'Sullivan, Ciara K; Prodromidis, Mamas I; Katakis, Ioanis
Departament: Enginyeria Química
Autor/s de la URV: Katakis, Ioanis / O'SULLIVAN, CIARA KATHLEEN
Paraules clau: Recombinase polymerase amplification Modified primers Karlodinium-veneficum Karlodinium armiger sequence in seawater Isothermal amplification Genomic dna Functionalized gold nanoparticles Electrochemical detection Dna detection Biosensors
Resum: A reagent-less DNA sensor has been developed exploiting a combination of gold nanoparticles, modified primers, and isothermal amplification. It is applied to the determination ofKarlodinium armiger, a toxic microalgae, as a model analyte to demonstrate this generic platform. Colloidal gold nanoparticles with an average diameter of 14 +/- 0.87 nm were modified with a mixed self-assembled monolayer of thiolated 33-mer DNA probes and (6-mercaptohexyl) ferrocene. Modified primers, exploiting a C3 spacer between the primer-binding site and an engineered single-stranded tail, were used in an isothermal recombinase polymerase amplification reaction to produce an amplicon by two single-stranded tails. These tails were designed to be complementary to a gold electrode tethered capture oligo probe, and an oligo probe immobilized on the gold nanoparticles, respectively. The time required for hybridization of the target tailed DNA with the surface immobilized probe and reporter probe immobilized on AuNPs was optimized and reduced to 10 min, in both cases. Amplification time was further optimized to be 40 min to ensure the maximum signal. Under optimal conditions, the limit of detection was found to be 1.6 fM of target dsDNA. Finally, the developed biosensor was successfully applied to the detection of genomic DNA extracted from a seawater sample that had been spiked with K. armiger cells. The demonstrated generic electrochemical genosensor can be exploited for the detection of any DNA sequence and ongoing work is moving towards an integrated system for use at the point-of-need.
Àrees temàtiques: Zootecnia / recursos pesqueiros Química Materiais Interdisciplinar Geociências Engenharias iv Engenharias ii Ciências biológicas i Ciências ambientais 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: ioanis.katakis@urv.cat
Identificador de l'autor: 0000-0003-4259-7098
Data d'alta del registre: 2024-10-26
Versió de l'article dipositat: info:eu-repo/semantics/acceptedVersion
Enllaç font original: https://link.springer.com/article/10.1007/s00604-021-04771-8
URL Document de llicència: https://repositori.urv.cat/ca/proteccio-de-dades/
Referència a l'article segons font original: Microchimica Acta. 188 (4): 117-
Referència de l'ítem segons les normes APA: AL-Madhagi, Sallam; O'Sullivan, Ciara K; Prodromidis, Mamas I; Katakis, Ioanis (2021). Combination of ferrocene decorated gold nanoparticles and engineered primers for the direct reagentless determination of isothermally amplified DNA. Microchimica Acta, 188(4), 117-. DOI: 10.1007/s00604-021-04771-8
DOI de l'article: 10.1007/s00604-021-04771-8
Entitat: Universitat Rovira i Virgili
Any de publicació de la revista: 2021
Tipus de publicació: Journal Publications