Rapid microfluidic solid-phase extraction system for hyper-methylated DNA enrichment and epigenetic analysis

被引:20
作者
De, Arpita [1 ]
Sparreboom, Wouter [1 ]
van den Berg, Albert [1 ]
Carlen, Edwin T. [1 ,2 ]
机构
[1] Univ Twente, MESA Inst Nanotechnol, BIOS Lab Chip Grp, NL-7522 NH Enschede, Netherlands
[2] Univ Tsukuba, Grad Sch Pure & Appl Sci, Tsukuba, Ibaraki 3058571, Japan
关键词
PCR; MICROCHIP; STOOL; PURIFICATION; ASSAYS;
D O I
10.1063/1.4899059
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 (mu SPE) 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 mu l elution volume and 92% efficiency using a 100 mu l elution volume. (C) 2014 AIP Publishing LLC.
引用
收藏
页数:11
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