Bubble-free rapid microfluidic PCR

Sang Hun Lee, Jihwan Song, Byungrae Cho, Soon Gweon Hong, Ori Hoxha, Taewook Kang, Dongchoul Kim, Luke P. Lee

Research output: Contribution to journalArticlepeer-review

55 Scopus citations

Abstract

Microfluidic polymerase chain reaction (PCR) has been of great interest owing to its ability to perform rapid and specific nucleic acid amplification and analysis on small volumes of samples. One of the major drawbacks of microfluidic PCR is bubble generation and reagent evaporation, which can cause malfunctions. Here, through theoretical modeling and characterization of bubble behavior, we propose a bubble-free microfluidic PCR device via controlled fluid transfer. Our approach exploits a thin impermeable polyethylene (PE) top layer that minimizes the generation of bubbles by inhibiting mass transport along a vertical direction. Simulation results demonstrate that a calculated mass flow difference of approximately 370% can be obtained by utilizing an impermeable membrane as the vertical barrier layer. To demonstrate proof-of-concept, two nanoporous polymeric materials, poly(dimethylsiloxane) (PDMS) and PE, were used for stand-alone self-powered sample loading (approximately 70 s) and for use as a vertical barrier layer, respectively. Consequently, we demonstrate successful amplification of the cMET gene, a nucleic acid (NA) biomarker for lung cancer, and complete an ultrafast PCR test in less than 3 min using a high powered Peltier-based thermal cycler under bubble-free conditions. This approach will result in a new paradigm for ultrafast molecular diagnosis and can facilitate NA-based nearly instantaneous diagnostics for point-of-care testing and for personalized and preventive medicine.

Original languageEnglish
Pages (from-to)725-733
Number of pages9
JournalBiosensors and Bioelectronics
Volume126
DOIs
StatePublished - 1 Feb 2019

Bibliographical note

Publisher Copyright:
© 2018

Keywords

  • Bubble-free microfluidic PCR
  • Lung cancer biomarker
  • Molecular diagnosis
  • Nanoporous polymers
  • Polymerase chain reaction

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