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Lookup NU author(s): Emeritus Professor Calum McNeilORCiD, Dr Neil Keegan
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
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.
Author(s): Jauset-Rubio M, Svobodová M, Mairal T, McNeil C, Keegan N, Saeed A, Abbas MN, El-Shahawi MS, Bashammakh AS, Alyoubi AO, O'Sullivan CK
Publication type: Article
Publication status: Published
Journal: Scientific Reports
Year: 2016
Volume: 6
Online publication date: 25/11/2016
Acceptance date: 31/10/2016
Date deposited: 25/11/2016
ISSN (electronic): 2045-2322
Publisher: Nature Publishing Group
URL: http://dx.doi.org/10.1038/srep37732
DOI: 10.1038/srep37732
Data Access Statement: http://dx.doi.org/10.17634/122638-2
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