Web of Science: 1 citas, Scopus: 1 citas, Google Scholar: citas,
In Situ Plant Virus Nucleic Acid Isothermal Amplification Detection on Gold Nanoparticle-Modified Electrodes
Khater, Mohga (Institut Català de Nanociència i Nanotecnologia)
De la Escosura Muñiz, Alfredo (Institut Català de Nanociència i Nanotecnologia)
Altet, Laura (Vetgenomics)
Merkoçi, Arben (Institut Català de Nanociència i Nanotecnologia)

Fecha: 2019
Resumen: Solid-phase isothermal recombinase polymerase amplification (RPA) offers many benefits over the standard RPA in homogeneous phase in terms of sensitivity, portability, and versatility. However, RPA devices reported to date are limited by the need for heating sources to reach sensitive detection. With the aim of overcoming such limitation, we propose here a label-free highly integrated in situ RPA amplification/detection approach at room temperature that takes advantage of the high sensitivity offered by gold nanoparticle (AuNP)-modified sensing substrates and electrochemical impedance spectroscopic (EIS) detection. Plant disease (Citrus tristeza virus (CTV)) diagnostics was selected as a relevant target for demonstration of the proof-of-concept. RPA assay for amplification of the P20 gene (387-bp) characteristic of CTV was first designed/optimized and tested by standard gel electrophoresis analysis. The optimized RPA conditions were then transferred to the AuNP-modified electrode surface, previously modified with a thiolated forward primer. The in situ-amplified CTV target was investigated by EIS in a Fe(CN ) /Fe(CN ) red-ox system, being able to quantitatively detect 1000 fg μL of nucleic acid. High selectivity against nonspecific gene sequences characteristic of potential interfering species such as Citrus psorosis virus (CPsV) and Citrus caxicia viroid (CCaV) was demonstrated. Good reproducibility (RSD of 8%) and long-term stability (up to 3 weeks) of the system were also obtained. Overall, with regard to sensitivity, cost, and portability, our approach exhibits better performance than RPA in homogeneous phase, also without the need of heating sources required in other solid-phase approaches.
Nota: Número d'acord de subvenció MINECO/MAT2017-87202-P
Nota: Número d'acord de subvenció MINECO/SEV-2013-0295
Derechos: Tots els drets reservats.
Lengua: Anglès.
Documento: article ; recerca ; acceptedVersion
Materia: Citrus tristeza virus ; Electrochemical impedance ; Gel electrophoresis analysis ; Gold nanoparticle-modified electrodes ; Interfering species ; Isothermal amplifications ; Long term stability ; Modified electrode surfaces
Publicado en: Analytical chemistry, Vol. 91, Issue 7 (April 2019) , p. 4790-4796, ISSN 1520-6882

DOI: 10.1021/acs.analchem.9b00340


Disponible a partir de: 2020-04-30
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El registro aparece en las colecciones:
Documentos de investigación > Documentos de los grupos de investigación de la UAB > Centros y grupos de investigación (producción científica) > Ciencias > Institut Català de Nanociència i Nanotecnologia (ICN2)
Artículos > Artículos de investigación
Artículos > Artículos publicados

 Registro creado el 2020-02-06, última modificación el 2020-03-25



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