68例矮小症患者的基因变异检测和病因研究

发布时间：2018-07-01 13:38

本文选题：矮小症 + 遗传病因　；参考：《北京协和医学院》2015年硕士论文

【摘要】：目的：矮小症的基因检测未在国内临床中广泛开展,很多单基因病尚未被诊断。且我国对矮小症的单基因病背景缺乏大规模的人群研究。本研究应用靶向基因捕获联合二代测序技术的方法对矮小症患者进行基因检测和病因诊断,旨在探究中国人群中病理性矮小症的单基因疾病谱,建立矮小症基因检测的方法并验证方法的有效性。方法：对门诊矮小患者进行临床评估和资料收集,纳入68例病因不明或可能与单基因突变相关的矮小患者,对其进行全血DNA提取,应用定制基因芯片对466个矮小相关基因的外显子进行捕获和二代测序,对变异进行筛选、解读和验证,发现致病基因突变和明确病因诊断。结果：68例矮小患者根据临床诊断分为五类,包括生长激素缺乏症20例,甲状腺功能减退症8例,骨软骨发育不良16例,综合症16例,特发性矮小症8例。1、总体检出结果：经变异检测、筛选和解读,在24例患者中检出23个基因突变,可能为患者的遗传病因,总检出率为35.3%。其中包括10例骨软骨发育不良、8例综合征、4例原发甲状腺功能低下、1例生长激素缺乏症和1例特发性矮小症。11个突变已经文献报道,12个突变未经报道。2、骨软骨发育不良疾病检测结果：变异检出率为62.5%,包括2例FGFR3突变、1例NPR2突变、2例COL2A1突变、3例FBN1突变、1例TRAPPC2突变和1例ARSE突变。其中6例有报道,4例未经文献报道。3、综合症检测结果：变异检出率为50%,包括5例单核苷酸变异和3例拷贝数变异。其中3例有文献报道,5例无文献报道。单核苷酸变异包括2例PTPN11突变、1例KRAS突变、1例NIPBL突变和1例PORCN突变。拷贝数变异分别为染色体17p13.3大片段杂合缺失、染色体4p16.3大片段杂合缺失和NFl基因2-15外显子的杂合重复。4、甲状腺功能减退症检测结果：变异检出率50%,包括2例NKX 2-1突变、1例NKX2-5-突变和1例PAX8突变。3例突变有文献报道,1例突变无报5、生长激素缺乏症检测结果：1例患者检出一条X染色体短臂杂合缺失和长臂杂合重复。6、特发性矮小症检测结果：1例患者检出位于染色体14q24.3的大片段杂合重复。结论：二代测序联合靶向基因捕获法能够解释35.3%的病理性矮小症的病因,它是一种准确、高效、低成本的基因变异检测方法,为临床矮小症的基因诊断和大规模研究提供了较好的途径。
[Abstract]:Objective: genetic detection of dwarfism has not been widely carried out in China, and many monogenic diseases have not been diagnosed. Moreover, there is a lack of large-scale population research on the background of monogenic disease of dwarfism in China. The aim of this study was to investigate the single gene disease spectrum of pathophysiological dwarfism in Chinese population by means of gene detection and etiological diagnosis by targeting gene capture combined with second-generation sequencing. To establish a method for the detection of dwarf gene and verify the effectiveness of the method. Methods: 68 dwarf patients with unknown etiology or possible single gene mutation were collected for clinical evaluation and data collection, and their whole blood DNA was extracted. The exons of 466 dwarfy-related genes were captured and sequenced with a customized gene chip. The mutation was screened, interpreted and verified, and the pathogenetic gene mutation was identified. Results according to the clinical diagnosis, 68 cases of short patients were divided into five categories, including 20 cases of growth hormone deficiency, 8 cases of hypothyroidism, 16 cases of osteochondral dysplasia and 16 cases of syndromes. There were 8 cases of idiopathic dwarfism. The result of total detection: 23 gene mutations were detected in 24 patients by mutation detection, screening and interpretation, which may be the genetic cause of the patients, the total detection rate was 35.3g. These include 10 cases of osteochondral dysplasia, 8 cases of syndromes, 4 cases of primary hypothyroidism, 1 case of growth hormone deficiency and 1 case of idiopathic dwarf. 11 mutations have been reported in the literature, 12 mutations have not been reported. The detection rate of abnormal diseases was 62.5%, including 2 cases of FGFR3 mutation and 1 case of NPR2 mutation, 2 cases of COL2A1 mutation, 3 cases of FBN1 mutation and 1 case of TRAPPC2 mutation and 1 case of ARSE mutation. In 6 cases, 4 cases were not reported in the literature. The result of syndrome detection was 50%, including 5 cases of single nucleotide mutation and 3 cases of copy number variation. Among them, 3 cases were reported in literature and 5 cases were not reported in literature. Single nucleotide mutation included 2 cases of PTPN11 mutation, 1 case of KRAS mutation and 1 case of NIPBL mutation and 1 case of PORCN mutation. Copy number variation was heterozygous deletion of large segment of chromosome 17p13.3. Chromosome 4p16.3 large fragment heterozygosity deletion and heterozygous repeat of exon 2-15 of NFL gene .4. results of hypothyroidism: the detection rate of mutation is 50, including 2 NKX2-1 mutations, 1 NKX2-5- mutation and 1 PAX8 mutation. 3 mutations have been reported. In this paper, we report the results of a case of growth hormone deficiency (GH deficiency) detected in 5 cases, and a short arm heterozygosity deletion of X chromosome and a long arm heterozygote repeat of .6 were detected in 1 case of growth hormone deficiency. The result of idiopathic dwarfism was that one patient was found to be located on chromosome 14q24.3. A large fragment heterozygote repeats. Conclusion: the combination of second-generation sequencing and targeted gene capture can explain 35.3% of the etiology of pathophysiological dwarfism. It is an accurate, efficient and low-cost method for detecting gene variation. It provides a good way for gene diagnosis and large-scale research of clinical dwarfism.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R725.8;R440

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