应用巢式PCR方法检测痉挛性斜颈多巴胺D5受体基因突变

发布时间：2018-03-02 12:46

本文选题：痉挛性斜颈　切入点：多巴胺D5受体　出处：《广西医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：背景：肌张力障碍的发生与脑内多巴胺、5-羟色胺及乙酰胆碱等神经递质的紊乱有关。痉挛性斜颈的发病可能与D5受体的基因多态性相关。对痉挛性斜颈患者D5受体基因微卫星多态性研究发现其等位基因4频率比对照组高，提示D5受体的突变可能是痉挛性斜颈的易感因素。D5受体的功能障碍可影响基底节多巴胺能通路，表明多巴胺受体功能异常与痉挛性斜颈的发病可能有相关性。前期研究中为探讨多巴胺D5受体基因多态性与痉挛性斜颈的相关性，发现了多巴胺D5受体基因（DRD5基因）的开放阅读框架ORF上877、1096、1199、1229、1268、1269位点出现单核苷酸位点的突变差异，其均有统计学意义，可能与痉挛性斜颈的易感性有关。目的：验证痉挛性斜颈前期研究中发现的DRD5基因中的ORF上877、1096、1199、1229、1268、1269位点的突变是否与其发病具有相关性，进一步为痉挛性斜颈的诊断提供分子生物学依据。方法：根据痉挛性斜颈患者前期研究中发现的ORF上877、1096、1199、1229、1268、1269位点出现单核苷酸位点的突变差异，在6个位点外围设计内、外侧引物，进行巢式PCR扩增。对巢式PCR扩增后得到目的基因进行直接基因测序，确定每个样本出现的基因点突变情况。结果：经过基因测序检测发现DRD5基因的ORF上877位点存在单核苷酸突变，由A变异为G，，与前期研究结果一致。ORF上1096、1199、1229、1268、1269位点无突变。ORF上877位点的单核苷酸突变经统计学分析得出p0.05具有统计学意义。该位点OR值为4.8331，多见于痉挛性斜颈组，提示该位点可能为痉挛性斜颈的危险因素。结论： 1.DRD5基因的ORF上1096、1199、1229、1268、1269位点无突变。 2.DRD5基因的ORF上877位点的单核苷酸位点的突变差异具有统计学意义，该位点可能与痉挛性斜颈有关，可能为痉挛性斜颈的危险因素。
[Abstract]:Background: dystonia is associated with the disorder of neurotransmitters such as dopamine 5-hydroxytryptamine and acetylcholine. The pathogenesis of spastic torticollis may be related to the gene polymorphism of D5 receptor. The frequency of allele 4 was higher than that of control group. These results suggest that the mutation of D5 receptor may be a susceptible factor of spasmodic torticollis. The dysfunction of D5 receptor may affect the dopaminergic pathway in basal ganglia. It is suggested that the abnormal function of dopamine receptor may be associated with the pathogenesis of spastic torticollis. In previous studies, the relationship between dopamine D5 receptor gene polymorphism and spasmodic torticollis was studied. It was found that the mutation of single nucleotide locus on the ORF of the open reading frame of dopamine D5 receptor gene (D5) was 877 / 10961 / 1999 / 1229 / 1268 / 1269, which was statistically significant and might be related to susceptibility to spasmodic torticollis. Objective: to investigate whether the mutation of ORF in the DRD5 gene in the prespasmodic study of spasmodic torticollis is related to the pathogenesis of spasmodic torticollis, and to provide molecular biological basis for the diagnosis of spasmodic torticollis. Methods: according to the mutation difference of single nucleotide locus on ORF, which was found in the previous study of spasmodic torticollis patients, there was a single nucleotide locus mutation at the locus 877, 1096, 1999, 1229, 1268, 1269, and the inner and outer primers were designed in the peripheral region of 6 loci. After nested PCR amplification, the target gene was sequenced directly to determine the point mutation of the gene in each sample. Results: a single nucleotide mutation was found in the ORF of DRD5 gene by gene sequencing. The mutation from A to G was consistent with the results of previous studies. The single nucleotide mutation at locus 109611991229C12681269 and locus 877 on ORF was statistically significant by statistical analysis. The OR value of the locus was 4.8331.It was more common in spasmodic torticollis group. These results suggest that this locus may be a risk factor for spasmodic torticollis. Conclusion:. 1. There was no mutation in the ORF of DRD5 gene. 2. The mutation of single nucleotide locus at locus 877 in ORF of DRD5 gene was statistically significant, which may be related to spasmodic torticollis and may be a risk factor for spasmodic torticollis.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R741

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