Rapid microfluidic solid-phase extraction system for hyper-methylated DNA enrichment and epigenetic analysis |
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| Authors: | Arpita De Wouter Sparreboom Albert van den Berg Edwin T Carlen |
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| Institution: | 1.BIOS Lab on a Chip Group, MESA+ Institute for Nanotechnology, University of Twente, Enschede 7522NH, The Netherlands;2.Graduate School of Pure and Applied Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8571, Japan |
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| Abstract: | Genetic sequence and hyper-methylation profile information from the promoter regions of tumor suppressor genes are important for cancer disease investigation. Since hyper-methylated DNA (hm-DNA) is typically present in ultra-low concentrations in biological samples, such as stool, urine, and saliva, sample enrichment and amplification is typically required before detection. We present a rapid microfluidic solid phase extraction (μSPE) system for the capture and elution of low concentrations of hm-DNA (≤1 ng ml−1), based on a protein-DNA capture surface, into small volumes using a passive microfluidic lab-on-a-chip platform. All assay steps have been qualitatively characterized using a real-time surface plasmon resonance (SPR) biosensor, and quantitatively characterized using fluorescence spectroscopy. The hm-DNA capture/elution process requires less than 5 min with an efficiency of 71% using a 25 μl elution volume and 92% efficiency using a 100 μl elution volume. |
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