应用高通量测序技术进行遗传性视网膜色素变性基因诊断的临床研究

发布时间：2018-01-11 11:18

本文关键词：应用高通量测序技术进行遗传性视网膜色素变性基因诊断的临床研究　出处：《昆明理工大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:遗传性视网膜色素变性(Retinitis Pigmentosa,RP)是一种遗传的眼科致盲性疾病,已报道与该病相关的致病基因有数十个,传统一代测序技术对这些致病基因的检测耗时长且检测通量低,而高通量测序(二代测序)能一次性高效的对数百个基因甚至人的全基因组外显子进行测序,因此被广泛运用于基因组学,由于目前高通量测序技术对RP临床基因诊断的运用尚少,故本论文课题将探究高通量测序技术对RP基因诊断的有效性、可行性、敏感性、特异性、安全性。方法:收集并提取四例RP患者外周血标本,定制RP相关致病基因的捕获探针,然后利用高通量测序技术对四例病例的外周血DNA样本进行RP相关致病基因的检测。结果:最终检测出:病例A的致病基因为USH2A,对应基因突变为c.4217CA和c.7068TG,进一步进行家系验证,初步得出病例A的RP致病突变为c.7068TG;病例B的致病基因为CRB1,对应基因突变为c.2172TA和c.3708TG;病例C的致病基因为RPE65,对应基因突变为c.1338GT;病例D的致病基因为RPE65,对应基因突变为c.1399CG和c.1338GT。结论:应用高通量测序技术对四例视网膜色素变性患者外周血标本进行RP相关致病基因检测和分析,均检出致病基因突变和疑似致病基因突变,而且发现了RP新的致病基因突变,因此本论文课题研究提示,应用高通量测序技术进行RP的基因诊断是准确和可行的,在临床上具有一定的实用价值。
[Abstract]:Objective: Retinitis Pigmentosa (RPP) is a genetic eye blindness disease, which has been reported to be associated with dozens of pathogenic genes. Traditional generation sequencing technology for the detection of these pathogenic genes takes a long time and low detection throughput, while high-throughput sequencing (second-generation sequencing) can effectively sequence hundreds of genes and even human genome exons. Therefore, it is widely used in genomics. Because of the lack of high-throughput sequencing in clinical gene diagnosis of RP, this paper will explore the effectiveness of high-throughput sequencing in the diagnosis of RP gene. Methods: the peripheral blood samples of four patients with RP were collected and extracted, and the capture probes of RP-related pathogenic genes were customized. RP-associated pathogenicity genes were detected by high-throughput sequencing in peripheral blood DNA samples of four cases. Results: finally, the pathogenic gene of case A was found to be USH2A. The corresponding gene mutations were c. 4217CA and c. 7068 TG. further pedigree validation showed that the RP-causing mutation of case A was c. 7068 TG. The pathogenicity gene of case B was CRB1, the corresponding gene mutation was c. 2172TA and c. 3708TG; The pathogenic gene of case C was RPE65 and the corresponding gene mutation was c. 1338 GTT. The pathogenic gene of case D was RPE65. The corresponding gene mutations were c.1399CG and c.1338GT.Conclusion: RP-related pathogenic genes were detected and analyzed in peripheral blood samples of four patients with retinitis pigmentosa by high-throughput sequencing. Both pathogenetic gene mutations and suspected pathogenetic gene mutations were detected, and new pathogenic gene mutations of RP were found. It is accurate and feasible to use high throughput sequencing technique to diagnose RP gene, and it has certain practical value in clinic.
【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R774.13;R440

【相似文献】