Development of High-Performance Gene Diagnosis System for Human Diseases Using Integrated Microdevice Technology

许多人类疾病是由致病基因的DNA序列的微小改变引起的。因此，人类疾病的DNA诊断是通过分析致病基因的DNA序列中的改变来实现的。在本研究中，我们开发了一种集成了PCR和毛细管阵列电泳的芯片微加工装置。通过对激光共聚焦荧光显微镜的改进，构建了用于该芯片的激光诱导荧光检测系统，并利用荧光显微镜研究了DNA的构象动力学。利用该系统可以检测到嵌入荧光染料标记的单个DNA分子在微通道阵列中的迁移，并观察了电场作用下单个DNA分子在线性聚丙烯酰胺填充的微通道中的构象动力学，以阐明DNA的分离机理，优化DNA分析的分离条件。在本研究中建立的最佳条件下，DNA片段高达1 kbp的分离时间仅为60 s。在这些情况下，这里显示的许多数据表明，集成微器件技术是一种很有前途的技术，用于人类疾病的DNA诊断，具有高通量，高精度，劳动密集度低，成本效益高。

Many human diseases are caused by small alterations in DNA sequence of the disease-causing genes. DNA diagnosis for human diseases is, therefore, realized by the analysis of the alterations in the DNA sequence of disease-causing genes. In this study, we developed microfabricated device integrated by PCR and capillary array electrophoresis on a chip. Laser-induced fluorescence detection system for the chip has been constructed by the modification of laser confocal fluorescence microscopy and we investigated DNA conformational dynamics using fluorescence microscopy. Even sigle DNA molecule labeled by intercalating fluorescence dyes was detected during migrating into the array of microchannels and conformational dynamics of single DNA molecule in microchannels filled with linear polyacrylamide under electric field was observed by this system to elucidate the separation mechanism of DNA and to optimize the separation conditions for DNA analysis. In the optimum conditions established in this study, DNA fragments up to 1 kbp were separated within only 60 s. In these instances, many data shown here demonstrate that integrated microdevice technology is a promising technology for DNA diagnosis of human diseases with high throughput, high accuracy, less labor-intensive, and cost effectiveness.