Articles producció científica> Enginyeria Química

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

  • Identification data

    Identifier: PC:1973
    Authors:
    Markéta SvobodováMiriam Jauset-RubioTeresa MairalCalum McNeilNeil KeeganMohammad S. El-ShahawiAbdulaziz S. BashammakhAbdulrahman O. AlyoubiCiara K. O´Sullivan
    Abstract:
    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).
  • Others:

    Author, as appears in the 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
    Department: Enginyeria Química
    URV's Author/s: 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
    Keywords: DNA Nucleic acids detection limits Lateral flow assay
    Abstract: 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).
    Research group: Group of Nanobiotechnology and Bioanalysis
    Thematic Areas: Chemical engineering Ingeniería química Enginyeria química
    licence for use: https://creativecommons.org/licenses/by/3.0/es/
    ISSN: 0003-2700
    Author identifier: n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; 0000-0002-0965-4655
    Record's date: 2016-11-23
    Last page: 10709
    Journal volume: 88
    Papper version: info:eu-repo/semantics/publishedVersion
    Link to the original source: https://pubs.acs.org/doi/10.1021/acs.analchem.6b03256
    Licence document URL: https://repositori.urv.cat/ca/proteccio-de-dades/
    Article's DOI: 10.1021/acs.analchem.6b03256
    Entity: Universitat Rovira i Virgili
    Journal publication year: 2016
    First page: 10701
    Publication Type: Article Artículo Article
  • Keywords:

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