A Simple, Low-Cost Platform for Real-Time Isothermal Nucleic Acid Amplification

Sensors (Basel). 2015 Sep 16;15(9):23418-30. doi: 10.3390/s150923418.

Abstract

Advances in microfluidics and the introduction of isothermal nucleic acid amplification assays have resulted in a range of solutions for nucleic acid amplification tests suited for point of care and field use. However, miniaturisation of instrumentation for such assays has not seen such rapid advances and fluorescence based assays still depend on complex, bulky and expensive optics such as fluorescence microscopes, photomultiplier tubes and sensitive lens assemblies. In this work we demonstrate a robust, low cost platform for isothermal nucleic acid amplification on a microfluidic device. Using easily obtainable materials and commercial off-the-shelf components, we show real time fluorescence detection using a low cost photodiode and operational amplifier without need for lenses. Temperature regulation on the device is achieved using a heater fabricated with standard printed circuit board fabrication methods. These facile construction methods allow fabrications at a cost compatible with widespread deployment to resource poor settings.

Keywords: helicase dependent amplification; isothermal amplification; molecular diagnostics; nucleic acid amplification test (NAAT); point-of care.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computer Systems / economics
  • DNA Helicases / metabolism
  • Equipment Design
  • Humans
  • Lab-On-A-Chip Devices / economics
  • Mobile Applications / economics
  • Molecular Diagnostic Techniques / instrumentation
  • Molecular Diagnostic Techniques / methods
  • Nucleic Acid Amplification Techniques / economics*
  • Nucleic Acid Amplification Techniques / instrumentation*
  • Nucleic Acid Amplification Techniques / methods
  • Point-of-Care Systems
  • Real-Time Polymerase Chain Reaction / economics
  • Real-Time Polymerase Chain Reaction / instrumentation
  • Temperature

Substances

  • DNA Helicases