聚二甲基硅氧烷微流控芯片电泳的临床应用研究

Clinical application research of polydimethylsiloxane microchip electrophoresis

目的探讨聚二甲基硅氧烷(PDMS)微流控芯片电泳技术的临床应用价值。方法利用模塑法制作PDMS微流控芯片,选择二胺基二苯甲烷(DDM)与羟丙基纤维素(HPC)对PDMS进行修饰,有效消除蛋白在芯片壁面上的吸附。PDMS微流控芯片电泳分离DNA标准品、PCR多重产物及血清脂蛋白。结果 10、20、50、100、200bp的DNA片段得到了很好的基线分离及良好重现性,峰面积的相对标准偏差分别为4.8%、6.3%、5.9%、3.5%、3.4%。线粒体疾病患者的多重PCR产物片段165、266、378、881bp在4min内实现了基线分离。血清低密度脂蛋白亚型、小而密低密度脂蛋白(sdLDL)实现了快速检测,冠心病组血清sdLDL检出率与健康体检组差异有统计学意义(P<0.01)。结论 PDMS微流控芯片电泳具有简单、快速、高效、低耗等特点,适合临床常规分析。

Objective To explore the clinical application of polydimethylsiloxane（PDMS） microchip electrophoresis.Methods The PDMS microchip was produced by molding a PDMS silicone elastomer against a microfabricated maser.Hydroxypropylcellulose（HPC） and n-Dodecyl β-D-maltoside（DDM） were utilized to alter channel surface to make it become hydrophilic and nonionic,thus reducing the interaction between the protein and the surface.DNA markers,multi-PCR products,and serum lipoproteins were separated by PDMS microchip electrophoresis.Results The reproducibility of the detection system was evaluated by Sybr Green labeled DNA markers containing five fragments,and relative standard deviation（RSD） value of peak area to each corresponding fragment could reach 4.8%,6.3%,5.9%,3.5% and 3.4% respectively.The multi-PCR products,including 165,266,378 and 881 bp fragments,could be successfully achieved baseline separation by this system within 4 min.The proportion of small,dense low density lipoprotein（sdLDL） were significantly higher in patients with coronary heart disease than in healthy controls（P0.01）.Conclusion PDMS microchip capillary electrophoresis could be a simple,rapid,effective and cost-low method,and could be a suitable analytical technique for clinical aplication.