Articles producció científica> Enginyeria Química

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

  • Datos identificativos

    Identificador: PC:2000
    Autores:
    Markéta SvobodováMiriam Jauset-RubioTeresa MairalCalum McNeilNeil KeeganAyman SaeedMohammad Nooredeen AbbasMohammad S. El-ShahawiAbdulaziz S. BashammakhAbdulrahman O. AlyoubiCiara K. O´Sullivan
    Resumen:
    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.
  • Otros:

    Autor según el artículo: 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
    Departamento: Enginyeria Química
    Autor/es 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
    Palabras clave: Metal Nanoparticles Nucleic Acid Amplification Techniques
    Resumen: 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.
    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: 2045-2322
    Identificador del 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
    Fecha de alta del registro: 2016-12-13
    Volumen de revista: 6
    Versión del articulo depositado: info:eu-repo/semantics/publishedVersion
    Enlace a la fuente original: https://www.nature.com/articles/srep37732
    URL Documento de licencia: https://repositori.urv.cat/ca/proteccio-de-dades/
    DOI del artículo: 10.1038/srep37732
    Entidad: Universitat Rovira i Virgili
    Año de publicación de la revista: 2016
    Página inicial: Art. num. 37732
    Tipo de publicación: Article Artículo Article
  • Palabras clave:

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