摘要
目的:通过聚合酶链式反应(PCR)、基因测序技术 及TDI FP技术,检测散发性与家族性帕金森病Parkin基因4 号外显子突变的差异性,探讨Parkin基因突变在PD发病机制 中的作用,同时建立一种新的突变检测方法.方法:以36例 家族性及114例散发性PD患者为实验对象,150名正常健康 人为阴性对照,通过PCR扩增含Parkin基因4号外显子的 DNA片段,电泳分析初次筛选片段缺失者.基因测序技术对 无缺失片段进行克隆、测序,筛选出突变型及野生型.最后应 用TDI FP技术检测R110或TAMRA标记ddNTP的FP值,鉴 定Parkin基因4号外显子的基因点突变型.结果:在36例家 族性PD患者中共发现10例4号外显子缺失突变,占27%,点 突变有4例,占11%;114例散发性PD患者中仅有2例4号 外显子缺失突变,占1%,而点突变则有48例,占42%.150例 阴性对照组中无缺失突变,点突变10例,占6%.
AIM: To study the difference of exon 4 mutation of Parkin gene between sporadic Parkinson's disease (SPD) and familial Parkinson's disease (FPD) by PCR, gene sequence detection and TDI-FP technique, to explore the important role of Parkin gene in the pathogenesis of Parkinson's disease, and to establish a new method for genotyping of mutation patterns. METHODS: The experiment group included 36 FPD patients and 114 SPD patients, and the control group consists of 150 healthy subjects. The target DNA fragment of Parkin gene exon 4 was amplified by polymerase chain reaction and the lost fragments were chosen. A mutation type and a wideness type were established by clone and gene sequence detection. The FP value of the R110 or TAMRA-labeled ddNTP was detected by TDI-FP assay to determine the SNP mutation genotype. RESULTS: Of the 36 cases with FPD, target DNA fragment lost was found in 10 cases (27%) and point gene mutation was found in 4 cases (11%). Of the 114 cases with SPD, target DNA fragment lost was found in only 2 (1%) cases, but point gene mutation were found in 48 (42%) cases. However, in the control group, no target DNA fragment lost was found, while point mutation was found in 10 (6%) cases. CONCLUSION: Parkin gene mutation may be one of the key factors in the pathogenesis of Parkinson's disease. Sporadic Parkinson's disease and familial Parkinson's disease have different mutation patterns, which may be related with the different pathogenesis of SPD and FPD.
出处
《第四军医大学学报》
北大核心
2005年第2期137-140,共4页
Journal of the Fourth Military Medical University
关键词
帕金森病
基因
突变
外显子
Parkinson disease
genes
mutation
exons
