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

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