Autor según el artículo: Markéta Svobodová; Miriam Jauset-Rubio; Teresa Mairal; Calum McNeil; Neil Keegan; Mohammad S. El-Shahawi; Abdulaziz S. Bashammakh; Abdulrahman O. Alyoubi; Ciara K. O´Sullivan

Departamento: Enginyeria Química

Autor/es de la URV: SVOBODOVÁ ., MARKÉTA; Miriam Jauset-Rubio; MAIRAL LERGA, TERESA; Calum McNeil; Neil Keegan; Mohammad S. El-Shahawi; Abdulaziz S. Bashammakh; Abdulrahman O. Alyoubi; O'SULLIVAN ., CIARA

Palabras clave: DNA Nucleic acids detection limits Lateral flow assay

Resumen: In this work, different methodologies were evaluated in search of robust, simple, rapid, ultrasensitive, and user-friendly lateral flow aptamer assays. In one approach, we developed a competitive based lateral flow aptamer assay, in which β-conglutin immobilized on the test line of a nitrocellulose membrane and β-conglutin in the test sample compete for binding to AuNP labeled aptamer. The control line exploits an immobilized DNA probe complementary to the labeled aptamer, forcing displacement of the aptamer from the β-conglutin-aptamer complex. In a second approach, the competition for aptamer binding takes place off-strip, and following competition, aptamer bound to the immobilized β-conglutin is eluted and used as a template for isothermal recombinase polymerase amplification, exploiting tailed primers, resulting in an amplicon of a duplex flanked by single stranded DNA tails. The amplicon is rapidly and quantitatively detected using a nucleic acid lateral flow with an immobilized capture probe and a gold nanoparticle labeled reporter probe. The competitive lateral flow is completed in just 5 min, achieving a detection limit of 55 pM (1.1 fmol), and the combined competitive-amplification lateral flow requires just 30 min, with a detection limit of 9 fM (0.17 amol).

Grupo de investigación: Group of Nanobiotechnology and Bioanalysis

Áreas temáticas: Chemical engineering Ingeniería química Enginyeria química

Acceso a la licencia de uso: https://creativecommons.org/licenses/by/3.0/es/

ISSN: 0003-2700

Identificador del autor: n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; 0000-0002-0965-4655

Fecha de alta del registro: 2016-11-23

Página final: 10709

Volumen de revista: 88

Versión del articulo depositado: info:eu-repo/semantics/publishedVersion

Enlace a la fuente original: https://pubs.acs.org/doi/10.1021/acs.analchem.6b03256

DOI del artículo: 10.1021/acs.analchem.6b03256

Entidad: Universitat Rovira i Virgili

Año de publicación de la revista: 2016

Página inicial: 10701

Tipo de publicación: Article Artículo Article