Reduced false positives and improved reporting of loop-mediated isothermal amplification using quenched fluorescent primers

Hardinge, Patrick ORCID: https://orcid.org/0000-0002-6292-5186 and Murray, James A. H. ORCID: https://orcid.org/0000-0002-2282-3839 2019. Reduced false positives and improved reporting of loop-mediated isothermal amplification using quenched fluorescent primers. Scientific Reports 9 , 7400. 10.1038/s41598-019-43817-z

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Abstract

Loop-mediated isothermal amplification (LAMP) is increasingly used in molecular diagnostics as an alternative to PCR based methods. There are numerous reported techniques to detect the LAMP amplification including turbidity, bioluminescence and intercalating fluorescent dyes. In this report we show that quenched fluorescent labels on various LAMP primers can be used to quantify and detect target DNA molecules down to single copy numbers. By selecting different fluorophores, this method can be simply multiplexed. Moreover this highly specific LAMP detection technique can reduce the incidence of false positives originating from mispriming events. Attribution of these events to particular primers will help inform and improve LAMP primer design.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Publisher:	Nature Publishing Group
ISSN:	2045-2322
Date of First Compliant Deposit:	2 May 2019
Date of Acceptance:	30 April 2019
Last Modified:	02 May 2023 20:16
URI:	https://orca.cardiff.ac.uk/id/eprint/122128

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