Development of DNA microarray-based reseqeuncing system for neurological diseases.

本项目的目的是开发一个高通量的神经系统疾病突变分析系统。该方法包括1.to开发微阵列基地reseqeuncing致病基因的神经系统疾病。2.散发性疾病候选基因综合分析的应用。3.综合突变分析基础上的表型变异分析。为了实现这些目标，我们开发了用于阿尔茨海默病、帕金森病、肌萎缩侧索硬化症、肾上腺脑白质营养不良、家族性痉挛性截瘫的DNA微阵列。我们已经证实，DNA微阵列碱基重测序的敏感性和特异性与基于双脱氧核苷酸链终止子的方法相当。我们应用DNA微阵列技术对43例痉挛性截瘫患者的致病基因进行了全面的突变分析。我们已经确定了7例SPG4患者和1例SPG3A患者。在SPG4患者中，有两例散发病例，表明突变的发生率降低。因此，我们已经证明了基于高通量DNA微阵列的再测序系统对于神经系统疾病的分子解剖是非常有用的，所述神经系统疾病包括阿尔茨海默氏病、帕金森病、肌萎缩性侧索硬化、肾上腺脑白质营养不良、家族性痉挛性截瘫、脊髓灰质炎和脊髓灰质炎。

The purpose of this project is to develop a high throughput mutational analysis system for neurological diseases. The approach consists of 1.to develop microarray bases reseqeuncing of causative genes for neurological diseases. 2.application of comprehensive analysis of candidate genes for sporadic diseases. and 3.analysis of phenotypic variations bases on comprehensive mutational analysis. To accomplish, these aims, we have developed DNA microarrays for Alzheimer's disease, Parkinson disease, amyotrophic lateral sclerosis, adrenoleukodsytrophy, familial spastic paraplegia. We have confirmed that the sensitivity and specificity of DNA microarray-bases reseqeuncing are comparable to those based on dideoxynucleotide chain terminator method. We have applied DMA microarray-based reseqeuncing for comprehensive mutational analysis of causative genes for 43 patients with spastic paraplegia. We have identified 7 patients with SPG4 and a patient with SPG3A. Among the SPG4 patients, there are two sporadic cases, suggesting reduced penetrance of the mutations. Thus, we have demonstrated that the high throughput DNA microarray-based resegeuncing system is highly useful for molecular dissection of neurological diseases including Alzheimer's disease, Parkinson disease, amyotrophic lateral sclerosis, adrenoleukodsytrophy, familial spastic paraplegia