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DNA序列的变化来实现的。在本研究中，我们开发了将PCR和毛细管阵列电泳集成在芯片上的微加工装置。通过对激光共聚焦荧光显微镜的改进，构建了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。该系统在电场作用下观察了单个DNA分子在填充线性聚丙烯酰胺微通道中的构象动力学，阐明了DNA的分离机理，优化了DNA分析的分离条件。在本研究建立的最佳条件下，在60秒内就能分离出1kbp的DNA片段。在这些情况下，这里显示的许多数据表明，集成微设备技术具有高通量、高精度、较少劳动密集型和成本效益，是一种很有前途的人类疾病DNA诊断技术。

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.