权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

DNA diagnosis by microchip capillary electrophoresis

微芯片毛细管电泳 DNA 诊断

基本信息

批准号：
12672252
负责人：
ARAKAWA Hidetoshi
金额：
$ 2.5万
依托单位：
Showa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12672252/
关键词：
micro chip electrophoresis microchip capillary electrophoresis vero toxin S. mutans S. sobrinus PCR う食細菌 Duplex PCR 遺伝子診断 S.mutanns S.sobrinus SSCP 腸管出血性大腸菌

项目摘要

Gene diagnosis is utilized as diagnoses such as hereditary disease and infectious disease, and as diagnoses method such as the prediction of drug metabolic ability. Generally the molecular biological technique utilizing in the laboratory is used as DNA analysis of gene diagnosis. However, the rapid and high sensitive DNA analysis has been required in infectious disease and tailor made medicine. Recently, as the method, DNA chip method is developed, and the use in the clinical field is being tried. In this study, microchip electrophoresis was developed as high sensitive and rapid DNA analytical method, and the establishment of gene diagnosis which analyze DNA polymorphism with the one point mutation, O157 ( intestinal hemorrhagic Escherichia coli ) and mutans streptococcus was carried out. Construction of micro chip and selection of the polymer which is used as the separation medium were carried out. As the result, it was possible to analyze the PCR product of the O157 verotoxin gene ( … More VT1 and VT2 ) using the channel of 30 μm depth and 100 μm width at separation length 1mm in slight10 seconds. And, in order to detect the one point mutation in the ras gene which is one of the oncogenes, the PCR was done mutation site, and next, SSCP (single-strand conformation polymorphism) analysis was carried out. In this study, separation media such as polymer and gel were mainly examined. As the result, the analysis of single nucleotide polymorphism (SNP) was difficult, though it was able to separate in the difference of large loop structure. Next, DNA diagnosis method of mutans streptococcus which existed in dental plaque, was developed as a diagnosis method of the dental treatment and care. Duplex allele specific PCR and common PCR for S. mutans and S. sobrinus were developed. And the obtained PCR products were analyzed by the imcrochip electrophoresis. By this method, these genes could be analyzed in 3 minutes. And, the detection limit was high sensitive of 20 fg. By the research of the above, the microchip electrophoresis seems to be the usefulness as a DNA detection method of the infectious disease which requires speedy DNA analysis. Less

基因诊断可用于遗传性疾病、感染性疾病等诊断，也可用于药物代谢能力预测等诊断方法。基因诊断中的DNA分析一般是实验室利用的分子生物学技术。然而，快速和高灵敏度的DNA分析已被要求在传染病和定制医学。最近，作为该方法，开发了DNA芯片方法，并且正在尝试在临床领域中使用。本研究将微芯片电泳技术作为一种高灵敏度、快速的DNA分析方法，建立了一点突变O157（肠出血性大肠杆菌）和变形链球菌DNA多态性分析的基因诊断方法。进行了微芯片的构建和作为分离介质的聚合物的选择。结果，可以分析O157 verotoxin基因的PCR产物（ ...更多信息 VT 1和VT 2），使用30 μm深、100 μm宽的通道，间隔长度为1mm，只需10秒。并且，为了检测癌基因之一的ras基因中的一个点突变，对突变位点进行PCR，然后进行SSCP（单链构象多态性）分析。在本研究中，主要考察了聚合物和凝胶等分离介质。因此，单核苷酸多态性（SNP）的分析是困难的，虽然它能够在大环结构的差异分离。其次，发展了牙菌斑中存在的变形链球菌的DNA诊断方法，作为牙科治疗和护理的诊断方法。等位基因特异性双重PCR和普通PCR。和S. sobrinus是发达国家。PCR产物经芯片电泳分析。通过这种方法，可以在3分钟内分析这些基因。检测限为20 fg。通过上述研究，微芯片电泳作为需要快速DNA分析的传染病的DNA检测方法似乎是有用的。少