A sample-to-answer droplet magnetofluidic assay platform for quantitative methylation-specific PCR

Biomed Microdevices. 2018 Mar 28;20(2):31. doi: 10.1007/s10544-018-0276-6.


Dysregulation of DNA methylation has been identified as an epigenetic biomarker for numerous cancer types. Gene-specific identification techniques relying on methylation-specific PCR (MSP) require a lengthy manual benchtop process that is susceptible to human-error and contamination. This MSP assay requires a series of discrete sample processing steps including genomic DNA extraction, bisulfite conversion and readout via PCR. In this work, we present a streamlined assay platform utilizing droplet magnetofluidic principles for integration of all sample processing steps required to obtain quantitative MSP signal from raw biological samples. We present a streamlined protocol for solid-phase extraction and bisulfite conversion of genomic DNA, which minimizes reagent use and simplifies the sample preparation protocol for implementation on a compact assay platform. Furthermore, we present a thermally robust assay chip that enables DNA extraction, bisulfite conversion and quantitative PCR from biological samples on a single device. Technical improvements to facilitate DNA extraction and PCR on a single chip in addition to chip performance characterization data are presented.

Keywords: Biomarkers; Epigenetics; Lab-on-chip; Magnetofluidics; Methylation; Methylation-specific PCR; qPCR.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • DNA Methylation*
  • Lab-On-A-Chip Devices*
  • Magnets*
  • Polymerase Chain Reaction / instrumentation*