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

Lookup NU author(s): Emeritus Professor Calum McNeil, Dr Neil Keegan

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


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.


Publication metadata

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 Source Location: http://dx.doi.org/10.17634/122638-2


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