FA mathematical model of recombinase polymerase amplification under continuously stirred conditions

Clint Moody, Heather Newell, Hendrik Viljoen

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Growing interest surrounds isothermal PCR techniques which have great potential for miniaturization for mobile diagnostics. Particularly promising, Recombinase Polymerase Amplification (RPA), combines this advantage of isothermal PCR with simplicity and rapid amplification. A mathematical model is presented of Recombinase Polymerase Amplification (RPA) and compared to experimental data. This model identifies the rate limiting steps in the chemical process, the effects of stirring, and insights in to using RPA for quantitative measurement of initial DNA concentration. Experiments are shown in which DNA amplification occurs under conditions of Couette flow and conditions of rotational turbulent flow. Hand mixing has been shown to dramatically shorten amplification times but introduces unpredictable variability. In some cases, this variability manifests itself as human error induced false negatives, a serious problem for all potential applications. Mechanical stirring demonstrates similarly short delay times while retaining high repeatability and reduces the potential for human error.

Original languageEnglish (US)
Pages (from-to)193-201
Number of pages9
JournalBiochemical Engineering Journal
Volume112
DOIs
StatePublished - Aug 15 2016

Keywords

  • Couette flow
  • Disease diagnostics
  • Isothermal PCR
  • Mathematical model
  • Recombinase Polymerase Amplification (RPA)

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Engineering
  • Bioengineering
  • Biomedical Engineering

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