BIOS Lab on a Chip Group, MESA+ Institute for Nanotechnology, University of Twente , Enschede 7522NH, The Netherlands.
Biomicrofluidics. 2014 Oct 21;8(5):054119. doi: 10.1063/1.4899059. eCollection 2014 Sep.
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.
肿瘤抑制基因启动子区域的遗传序列和超甲基化谱信息对于癌症疾病的研究非常重要。由于超甲基化 DNA(hm-DNA)在粪便、尿液和唾液等生物样本中的浓度通常极低,因此在检测前通常需要进行样本富集和扩增。我们提出了一种基于蛋白质-DNA 捕获表面的快速微流固相萃取(μSPE)系统,用于从小体积中捕获和洗脱低浓度的 hm-DNA(≤1ng/ml),使用被动微流控芯片平台。使用实时表面等离子体共振(SPR)生物传感器对所有测定步骤进行了定性表征,并使用荧光光谱法进行了定量表征。hm-DNA 的捕获/洗脱过程耗时不到 5 分钟,使用 25μl 洗脱体积的效率为 71%,使用 100μl 洗脱体积的效率为 92%。