精神分裂症易感基因突变检测

发布时间：2018-05-11 00:44

本文选题：精神分裂症 + DISC1　；参考：《济南大学》2013年硕士论文

【摘要】：精神分裂症（Schizophrenia，MIM181500）是一种以思维、情感、行为异常，精神活动与环境不协调为特征的严重精神障碍，多发病于青壮年。在世界人口中的终生患病率为3.8‰-8.4‰；占全球范围内15～44岁人群疾病负担的2.6%，是世界上导致残疾的第四大原因。精神分裂症的病因仍未查明，目前普遍认为遗传因素对其病因有重大影响，因为精神分裂症的遗传度约为80%，患者的一级亲属发病风险是普通人的5到10倍。精神分裂症的遗传机制十分复杂，一般认为是由多对微效基因与环境因素共同作用所导致。通过遗传连锁和关联分析，人们发现了很多与精神分裂症病因有关的染色体区域和候选基因。精神分裂症断裂基因（DISRUPTED IN SCHIZOPHRENIA， DISC1）位于1q42.2；全长约410kb，包含13个外显子，已发现有16个转录本。此基因编码854个氨基酸的蛋白质，包括一个富含丝氨酸、丙氨酸、甘氨酸的球状N端区和一个由3-13号外显子编码形成的卷曲的C端区。羧基端区域富含许多环状结构域，有利于DISC1与其他蛋白间的结合。锌指蛋白804A（ZINC FINGER PROTEIN804A， ZNF804A）位于2q31，2008年，O’DONOVAN等人在全世界范围内首次发现了基于GWAS的精神分裂症风险因子——遗传标记rs1344706，并鉴定出首选致病基因ZNF804A。随后来自其他3个研究小组独立的GWAS研究均证实了rs1344706与精神分裂症的关联，从而进一步证明ZNF804A的研究价值。目的对精神分裂症患者的DISC1基因和ZNF804A基因的全部外显子进行突变筛查，以求发现突变。方法阶段一：DISC1基因外显子突变筛查选择了2004年8月在山东省济南市精神卫生防治中心与济南市历城区锦绣川精神病疗养院住院的精神分裂症患者96例；正常对照者96例取自山东省血液中心献血者。对DISC1基因的13个外显子合成26对引物，，对96例精神分裂症患者与96例正常对照者的DNA样本分别进行聚合酶链扩增后，通过高分辨率熔解曲线（High Resolution Melting， HRM）方法对扩增产物进行突变检测。阶段二：ZNF804A基因外显子突变筛查选择了2004年8月在山东省济南市精神卫生防治中心与济南市历城区锦绣川精神病疗养院住院的精神分裂症患者182例；正常对照者96例取自山东省血液中心献血者。对DISC1基因的4个外显子合成20对引物，将182例精神分裂症患者的DNA构建2个DNA混合池，将96例正常对照者的DNA样本构建一个DNA混合池。首先将上述3个DNA混合池分别进行聚合酶链扩增，然后再将稀释后的扩增产物进行COLD-PCR聚合酶链二次扩增，最后用高分辨率熔解曲线（High Resolution Melting， HRM）方法对扩增产物进行突变检测。结果 1.用HRM方法对96例精神分裂症患者和96例正常对照者的DISC1基因外显子扩增产物的熔解曲线分析未发现两者的熔解曲线存在差别。 2.用HRM方法对182例精神分裂症患者和96例正常对照者的ZNF804A基因外显子扩增产物熔解曲线分析在精神分裂症患者Exon4.6位点发现多态性。结论本实验结果表明，在精神分裂症患者DISC1基因外显子的突变筛查没有发现突变，在精神分裂症患者ZNF804A基因Exon4.6位点发现多态性。DISC1基因可能与山东省精神分裂症患者的发病无关，ZNF804A基因与山东精神分裂症患者发病的关系仍待进一步研究。
[Abstract]:Schizophrenia (MIM181500) is a serious mental disorder characterized by thought, emotion, abnormal behavior and incoordination between mental activity and environment. It is mostly in young adults. The lifetime prevalence rate in the world population is 3.8 per thousand -8.4 per thousand; it accounts for 2.6% of the disease burden of 15~44 years old in the world, which is the cause of disability in the world. Fourth major causes. The cause of schizophrenia is still not identified. It is widely believed that genetic factors have a significant impact on the cause of schizophrenia, because the heritability of schizophrenia is about 80%, the risk of first-degree relatives of the patients is 5 to 10 times that of ordinary people. The genetic mechanism of schizophrenia is very complex, generally thought to be from multiple pairs of genes and rings. Genetic linkage and association analysis have revealed many chromosomal regions and candidate genes related to the etiology of schizophrenia.
The schizophrenia gene (DISRUPTED IN SCHIZOPHRENIA, DISC1) is located in 1q42.2; a total of about 410KB, containing 13 exons and 16 transcripts. This gene encodes a 854 amino acid protein, including a spherical N endpoint rich in serine, alanine, glycine, and a curly C encoded by exon 3-13. The terminal region is rich in many ring domains, which is beneficial to the binding of DISC1 to other proteins.
The zinc finger protein 804A (ZINC FINGER PROTEIN804A, ZNF804A) was located in 2q312008, and O 'DONOVAN and others first discovered the risk factor of schizophrenia based on GWAS - genetic marker rs1344706, and identified the preferred pathogenic gene ZNF804A. subsequently from the independent GWAS studies of the other 3 research groups. 706 the association between schizophrenia and schizophrenia, thus further proving the value of ZNF804A research.
objective
Mutation screening of all exons of DISC1 gene and ZNF804A gene was performed in schizophrenic patients in order to detect mutations.
Method
Stage 1: DISC1 gene exon mutation screening
96 schizophrenic patients were selected in August 2004 in Ji'nan city of Shandong mental health prevention and control center and Ji'nan City District Jinxiu Sichuan spiritual sanatorium. 96 cases from the blood center of Shandong province were taken from the blood center of Shandong province. 13 exons of DISC1 gene were synthesized in 26 pairs, 96 schizophrenic patients and 96 cases were positive. The DNA samples of the normal controls were amplified by polymerase chain reaction, and the amplified products were detected by the high resolution fusion curve (High Resolution Melting, HRM).
Stage two: exon mutation screening for ZNF804A gene
182 schizophrenic patients were selected in August 2004 in Ji'nan city of Shandong mental health prevention and control center and Ji'nan City District Jinxiu Sichuan spiritual sanatorium. 96 cases from the blood center of Shandong province were taken from the blood center of Shandong province. 4 exons of DISC1 gene were synthesized in 20 pairs, and 182 schizophrenic patients were constructed. 2 DNA mixed pools were used to construct a DNA mixing pool in the DNA samples of 96 normal controls. First, the 3 DNA mixed pools were amplified by polymerase chain reaction, and then the amplified products were amplified by COLD-PCR polymerase chain for two times. Finally, the high resolution melting curve (High Resolution Melting, HRM) method was used to amplify the product. Mutation detection.
Result
1. the fusion curves of DISC1 exon amplification products of 96 schizophrenic patients and 96 normal controls were analyzed by HRM method, and there was no difference in the melting curve between the two.
2. HRM method was used to analyze the fusion curve of the exon amplification product of ZNF804A gene exon in 182 schizophrenic patients and 96 normal controls. The polymorphism was found in the Exon4.6 locus of schizophrenic patients.
conclusion
The results of this study showed that mutation screening was not found in the DISC1 exons of the schizophrenic patients. The polymorphism of the.DISC1 gene in the ZNF804A gene Exon4.6 locus in schizophrenic patients was not found to be related to the incidence of schizophrenia in Shandong, and the relationship between the ZNF804A gene and the incidence of Shandong schizophrenic patients was still to be entered. One step of research.

【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.3

【参考文献】