Articles producció científica> Enginyeria Química

Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay

  • Dades identificatives

    Identificador: PC:2000
    Autors:
    Markéta SvobodováMiriam Jauset-RubioTeresa MairalCalum McNeilNeil KeeganAyman SaeedMohammad Nooredeen AbbasMohammad S. El-ShahawiAbdulaziz S. BashammakhAbdulrahman O. AlyoubiCiara K. O´Sullivan
    Resum:
    Sensitive, specific, rapid, inexpensive and easy-to-use nucleic acid tests for use at the point-of-need are critical for the emerging field of personalised medicine for which companion diagnostics are essential, as well as for application in low resource settings. Here we report on the development of a point-of-care nucleic acid lateral flow test for the direct detection of isothermally amplified DNA. The recombinase polymerase amplification method is modified slightly to use tailed primers, resulting in an amplicon with a duplex flanked by two single stranded DNA tails. This tailed amplicon facilitates detection via hybridisation to a surface immobilised oligonucleotide capture probe and a gold nanoparticle labelled reporter probe. A detection limit of 1 × 10-11 M (190 amol), equivalent to 8.67 × 105 copies of DNA was achieved, with the entire assay, both amplification and detection, being completed in less than 15 minutes at a constant temperature of 37 °C. The use of the tailed primers obviates the need for hapten labelling and consequent use of capture and reporter antibodies, whilst also avoiding the need for any post-amplification processing for the generation of single stranded DNA, thus presenting an assay that can facilely find application at the point of need.
  • Altres:

    Autor segons l'article: Markéta Svobodová; Miriam Jauset-Rubio; Teresa Mairal; Calum McNeil; Neil Keegan; Ayman Saeed; Mohammad Nooredeen Abbas; 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; Ayman Saeed; Mohammad Nooredeen Abbas; Mohammad S. El-Shahawi; Abdulaziz S. Bashammakh; Abdulrahman O. Alyoubi; O'SULLIVAN, CIARA
    Paraules clau: Metal Nanoparticles Nucleic Acid Amplification Techniques
    Resum: Sensitive, specific, rapid, inexpensive and easy-to-use nucleic acid tests for use at the point-of-need are critical for the emerging field of personalised medicine for which companion diagnostics are essential, as well as for application in low resource settings. Here we report on the development of a point-of-care nucleic acid lateral flow test for the direct detection of isothermally amplified DNA. The recombinase polymerase amplification method is modified slightly to use tailed primers, resulting in an amplicon with a duplex flanked by two single stranded DNA tails. This tailed amplicon facilitates detection via hybridisation to a surface immobilised oligonucleotide capture probe and a gold nanoparticle labelled reporter probe. A detection limit of 1 × 10-11 M (190 amol), equivalent to 8.67 × 105 copies of DNA was achieved, with the entire assay, both amplification and detection, being completed in less than 15 minutes at a constant temperature of 37 °C. The use of the tailed primers obviates the need for hapten labelling and consequent use of capture and reporter antibodies, whilst also avoiding the need for any post-amplification processing for the generation of single stranded DNA, thus presenting an assay that can facilely find application at the point of need.
    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: 2045-2322
    Identificador de l'autor: n/a; n/a; n/a; n/a; n/a; 0000-0001-5760-5180; 0000-0002-7142-7322; n/a; n/a; n/a; 0000-0002-0965-4655
    Data d'alta del registre: 2016-12-13
    Volum de revista: 6
    Versió de l'article dipositat: info:eu-repo/semantics/publishedVersion
    Enllaç font original: https://www.nature.com/articles/srep37732
    URL Document de llicència: https://repositori.urv.cat/ca/proteccio-de-dades/
    DOI de l'article: 10.1038/srep37732
    Entitat: Universitat Rovira i Virgili
    Any de publicació de la revista: 2016
    Pàgina inicial: Art. num. 37732
    Tipus de publicació: Article Artículo Article
  • Paraules clau:

    Detectors químics
    ADN
    Metal Nanoparticles
    Nucleic Acid Amplification Techniques
    Chemical engineering
    Ingeniería química
    Enginyeria química
    2045-2322
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