Author, as appears in the 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
Department: Enginyeria Química
URV's Author/s: 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
Keywords: Metal Nanoparticles Nucleic Acid Amplification Techniques
Abstract: 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.
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: 2045-2322
Author identifier: 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
Record's date: 2016-12-13
Journal volume: 6
Papper version: info:eu-repo/semantics/publishedVersion
Link to the original source: https://www.nature.com/articles/srep37732
Article's DOI: 10.1038/srep37732
Entity: Universitat Rovira i Virgili
Journal publication year: 2016
First page: Art. num. 37732
Publication Type: Article Artículo Article
Repositori URV