Real-time duplex applications of loop-mediated AMPlification (LAMP) by assimilating probes

International Journal of Molecular Sciences
R Kubota, Daniel M Jenkins

Abstract

Isothermal nucleic-acid amplification methods such as Loop-Mediated isothermal AMPlification (LAMP) are increasingly appealing alternatives to PCR for use in portable diagnostic system due to the low cost, weight, and power requirements of the instrumentation. As such, interest in developing new probes and other functionality based on the LAMP reaction has been intense. Here, we report on the development of duplexed LAMP assays for pathogen detection using spectrally unique Assimilating Probes. As proof of principle, we used a reaction for Salmonella enterica as a model coupled with a reaction for λ-phage DNA as an internal control, as well as a duplexed assay to sub-type specific quarantine strains of the bacterial wilt pathogen Ralstonia solanacearum. Detection limits for bacterial DNA analyzed in individual reactions was less than 100 genomic equivalents in all cases, and increased by one to two orders of magnitude when reactions were coupled in duplexed formats. Even so, due to the more robust activity of newly available strand-displacing polymerases, the duplexed assays reported here were more powerful than analogous individual reactions reported only a few years ago, and represent a significant advance for incorporation o...Continue Reading

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Citations

Aug 1, 2020·PLoS Neglected Tropical Diseases·Laura C BonneyRoger Hewson
Sep 10, 2020·Journal of Plant Pathology : an International Journal of the Italian Phytopathological Society·Jugpreet SinghAwais Khan
May 1, 2021·International Journal of Environmental Research and Public Health·Yufei ChenFengjie Sun
May 21, 2021·MSphere·Sanchita BhadraAndrew D Ellington

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Datasets Mentioned

BETA
GMI1000

Methods Mentioned

BETA
nucleic acid amplification
Fluorescence
PCR

Software Mentioned

PrimerExplorer

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