Elfimova Natalia, Amer Wafa, Odenthal Margarete
Institute for Pathology, University of Cologne, Cologne, Germany.
Methods Mol Biol. 2013;919:287-96. doi: 10.1007/978-1-62703-029-8_25.
Microsatellites are short repetitive sequences of two, three, or four bases, prone to base mispairing. Microsatellite instability (MSI) occurs frequently in various types of cancer due to a defective DNA mismatch repair system. Therefore, MSI analysis is an important tool in clinical research and molecular diagnostics. Mostly, polyacrylamide gel electrophoresis or capillary electrophoresis of labeled microsatellite sequences is used for the detection of MSI. Here we present a microfluidic-based electrophoresis technology for MSI analyses. Defined loci of microsatellites were PCR amplified and amplicons were analyzed by microfluidic-based electrophoresis. The electropherogram profiles of tumor and non-tumor derived DNA clearly revealed an individual pattern identifying differences in tumor-associated microsatellites. Detection of MSI by microfluidics turned out to be a simple and efficient procedure but less laborious than conventional approaches. Thus, the chip-based microfluidic electrophoresis is a simple, reliable, and robust technology for MSI detection, which allows label-free analyses of microsatellite amplicons within 30 min.
微卫星是由两个、三个或四个碱基组成的短重复序列,容易发生碱基错配。由于DNA错配修复系统存在缺陷,微卫星不稳定性(MSI)在各种类型的癌症中频繁出现。因此,MSI分析是临床研究和分子诊断中的一项重要工具。大多数情况下,标记微卫星序列的聚丙烯酰胺凝胶电泳或毛细管电泳用于MSI检测。在此,我们介绍一种基于微流控的电泳技术用于MSI分析。对微卫星的特定基因座进行PCR扩增,并通过基于微流控的电泳对扩增产物进行分析。肿瘤和非肿瘤来源DNA的电泳图谱清晰地显示出一种独特模式,可识别肿瘤相关微卫星的差异。事实证明,通过微流控检测MSI是一个简单高效的过程,且比传统方法省力。因此,基于芯片的微流控电泳是一种用于MSI检测的简单、可靠且稳健的技术,它能够在30分钟内对微卫星扩增产物进行无标记分析。