Development of Integrated Micro-Chip Device for DNA Analysis

DNA芯片系统的开发，可以在小型医疗机构中进行高精度的个人遗传信息的即时分析，而无需专门技能，可以使医生能够进行早期治疗，并通过其准确的发育前诊断防止其进一步发展。本研究的目的是开发高精度的微积累DNA芯片，用于基因分析，以准确地进行疾病发生前的诊断。DNA芯片由扩增待分析DNA的PCR和使用杂交的分析部分组成。今年，我们开发了微型PCR设备和新的DNA分析技术。我们通过在3cm×3cm的玻璃板上加工微波路径，制作了温度循环约30次的微型PCR设备。还制作了微型加热器， ...更多信息 通过在已经加工了波路的玻璃板的背面上形成ITO膜来控制温度。通过将芘酸溶液送入微波通道并测量显微荧光，分析了微通道内的温度分布，结果表明ITO膜可有效地作为PCR芯片的微加热器。此外，通过以大肠杆菌为模板进行DNA的PCR，确认了国产PCR探针的扩增能力与市场上的PCR探针一样。此外，开发了使用胶体金的DNA杂交分析新技术。将边缘修饰有金胶体的DNA固定在金层上，并利用表面等离子体共振进行光谱测量。因此，本实验使我们能够观察到大的SPR信号伴随着杂交。通过本研究方法对检测限的进一步检验，证实了pM水平检测的有效性。本年度的研究成果是建立了微量PCR创造技术和成功开发了新的DNA杂交检测技术。少

The development of DNA chip system, which can carry out immediate analysis on an individual genetic information with high accuracy without specialized skill in minor medical institutions, could enable a doctor to perform the early treatment and prevent it from further developing with its accurate pre-development diagnosis. This is expected to contribute greatly to the disease prevention and maintenance of health conditions of a people.Our research aims at the development of high accuracy micro-accumulation DNA chip, which can be applicable to a gene analysis for an accurate pre-disease development diagnosis. DNA chip consists of PCR, which amplifies DNA to be analyzed, and analyzing portion, which uses hybridization. This year, we have developed micro-PCR equipment and new DNA analysis teqnique.We nave created micro PCR equipment whose temperature cycles are about as many as 30 times by processing the microwave route on the glass board of 3cm×3cm Micro-heater has also been created for … More temperature control by forming ITO film on the reverse side of the glass board on which a wave route has been processed. Analysis on the temperature profile in the micro channel by sending pyrenes acid solution into the microwave route and by measuring microscopic fluorescence has revealed that ITO film is effectively functioned as the micro heater of PCR chip. Also, carrying out PCR of DNA whose origin is a colon bacillus as template has made us confirm that the home-developed PCR tip can be amplified as much as the one on the market.Also, we have developed new DNA hybridization analysis technique using gold colloid. The prove DNA decorated with the gold colloid at the edge has been fixed on the gold layer and spectrum measurement using surface plasmon resonance has been carried out. This experiment consequently has enabled us to observe large SPR signal accompanied by hybridization. Further examination of detection limit by our research method makes us confirm the availability of pM level-detection. Furthermore, the effectiveness for SNPs detection has also been confirmed ; therefore, this experimental method is proved to be applicable to DNA detection.Our research achievements of this year are the establishment of micro PCR creation tequnique and the successful development of new DNA hybridization detection technique. Less