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

  • Dades identificatives

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

    Autor segons l'article: 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
    Departament: Enginyeria Química
    Autor/s 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
    Paraules clau: DNA Nucleic acids detection limits Lateral flow assay
    Resum: 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).
    Grup de recerca: Group of Nanobiotechnology and Bioanalysis
    Àrees temàtiques: Chemical engineering Ingeniería química Enginyeria química
    Accès a la llicència d'ús: https://creativecommons.org/licenses/by/3.0/es/
    ISSN: 0003-2700
    Identificador de l'autor: n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; 0000-0002-0965-4655
    Data d'alta del registre: 2016-11-23
    Pàgina final: 10709
    Volum de revista: 88
    Versió de l'article dipositat: info:eu-repo/semantics/publishedVersion
    Enllaç font original: https://pubs.acs.org/doi/10.1021/acs.analchem.6b03256
    URL Document de llicència: https://repositori.urv.cat/ca/proteccio-de-dades/
    DOI de l'article: 10.1021/acs.analchem.6b03256
    Entitat: Universitat Rovira i Virgili
    Any de publicació de la revista: 2016
    Pàgina inicial: 10701
    Tipus de publicació: Article Artículo Article
  • Paraules clau:

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