10 Citations (Scopus)

Abstract

This work describes an enhancement to the loop-mediated isothermal amplification (LAMP) reaction which results in improved performance. Enhancement is achieved by adding a new set of primers to conventional LAMP reactions. These primers are termed "swarm primers" based on their relatively high concentration and their ability to create new amplicons despite the theoretical lack of single-stranded annealing sites. The primers target a region upstream of the FIP/BIP primer recognition sequences on opposite strands, substantially overlapping F1/B1 sites. Thus, despite the addition of a new primer set to an already complex assay, no significant increase in assay complexity is incurred. Swarm priming is presented for three DNA templates: Lambda phage, Synechocystis sp. PCC 6803 rbcL gene, and human HFE. The results of adding swarm primers to conventional LAMP reactions include increased amplification speed, increased indicator contrast, and increased reaction products. For at least one template, minor improvements in assay repeatability are also shown. In addition, swarm priming is shown to be effective at increasing the reaction speed for RNA amplification via RT-LAMP. Collectively, these results suggest that the addition of swarm primers will likely benefit most if not all existing LAMP assays based on state-of-the-art, six-primer reactions.

Original languageEnglish (US)
Pages (from-to)625-632
Number of pages8
JournalAnalytical Chemistry
Volume89
Issue number1
DOIs
StatePublished - Jan 3 2017

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Amplification
Assays
Speed indicators
Bacteriophages
Reaction products
Genes
RNA
Annealing
DNA

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Improved Performance of Loop-Mediated Isothermal Amplification Assays via Swarm Priming. / Martineau, Rhett L.; Murray, Sarah A.; Ci, Shufang; Gao, Weimin; Chao, Shih-Hui; Meldrum, Deirdre.

In: Analytical Chemistry, Vol. 89, No. 1, 03.01.2017, p. 625-632.

Research output: Contribution to journalArticle

Martineau, Rhett L. ; Murray, Sarah A. ; Ci, Shufang ; Gao, Weimin ; Chao, Shih-Hui ; Meldrum, Deirdre. / Improved Performance of Loop-Mediated Isothermal Amplification Assays via Swarm Priming. In: Analytical Chemistry. 2017 ; Vol. 89, No. 1. pp. 625-632.
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