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Aptamer Lateral Flow Assays for Ultrasensitive Detection of β-Conglutin Combining Recombinase Polymerase Amplification and Tailed Primers

  • Datos identificativos

    Identificador: PC:1973
    Autores:
    Markéta SvobodováMiriam Jauset-RubioTeresa MairalCalum McNeilNeil KeeganMohammad S. El-ShahawiAbdulaziz S. BashammakhAbdulrahman O. AlyoubiCiara K. O´Sullivan
    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).
  • Otros:

    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
    URL Documento de licencia: https://repositori.urv.cat/ca/proteccio-de-dades/
    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
  • Palabras clave:

    Biotecnologia
    Àcids nucleics -- Anàlisi
    DNA
    Nucleic acids
    detection limits
    Lateral flow assay
    Chemical engineering
    Ingeniería química
    Enginyeria química
    0003-2700
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