全基因组芯片技术在先天性心脏病基因组拷贝数变异中的研究

发布时间：2018-04-25 21:03

本文选题：先天性心脏病 + 拷贝数变异　；参考：《浙江大学》2014年硕士论文

【摘要】：目的：采用全基因组芯片检测技术对先天性心脏病患儿进行全基因组DNA检测,精细定位研究DNA的致病性拷贝数变异,比较DNA拷贝数突变与先天性心脏病发病及临床表型之间的关系,探讨全基因组芯片检测技术在先天性心脏病患儿临床诊断中的可行性。对象与方法：选取35例心脏B超及手术确诊的散发先天性心脏病患儿,按照无菌要求、EDTA抗凝,采集外周血2-4ml,应用Gentra Puregene Kit试剂盒(德国Qiagen GMBH公司)分别提取基因组DNA,将基因组DNA与Affymetrix Genome-Wide Human SNP Array6.0芯片杂交后进行扫描,得到原始数据后再用CHAS(Chromosome Analysis Suite)软件分析,结合文献得出结果。结果：在35例先天性心脏病患儿中共检出6例染色体DNA异常,检出率为17.14%,基因芯片结果显示：①病例1存在22q11.21缺失合并22q11.1重复,分别位于16055170bp-17023514bp之间和18884837bp-21465481bp之间,诊断为22q11缺失综合征合并猫眼综合征；②病例2为47,XY,+21,结合患儿染色体核型分析结果,诊断为21-三体综合征；③病例3存在22q11.21缺失,位于18793934bp-21431554bp之间,诊断为22q11缺失综合征；④病例4存在22q11.21缺失,位于20716876bp-21800471bp之间,为22q11的不典型缺失；⑤病例5为47,XXX,诊断为47,XXX综合征；⑥病例6存在Yp11.31-q11.221重复,位于2650424bp-16107288bp之间和Yq11.22q11.23缺失,位于16167646bp-28799654bp之间,为ZAF(无精子因子)缺失。结论：与传统的细胞遗传分析方法相比,全基因组芯片扫描技术能够高分辨、高通量和高准确性地检测出亚显微水平的染色体畸变,有利于病理性拷贝数变异和致病基因的筛查,有利于核型-表型的相关性研究,可以作为传统的细胞遗传分析方法的有益补充应用于临床细胞遗传诊断中。
[Abstract]:Objective: The whole genome DNA of children with congenital heart disease was detected by whole genome microarray technique. The pathogenicity copy number variation of DNA was studied by fine localization, and the relationship between DNA copy number mutation and the pathogenesis and clinical phenotype of congenital heart disease was compared. To explore the feasibility of whole-genome microarray in the diagnosis of children with congenital heart disease. Objects and methods: 35 children with congenital heart disease diagnosed by echocardiography and surgical diagnosis were selected and treated with EDTA for anticoagulation according to aseptic requirements. Peripheral blood 2-4ml was collected, genomic DNA was extracted by Gentra Puregene Kit kit (Qiagen GMBH Company, Germany), genomic DNA was hybridized with Affymetrix Genome-Wide Human SNP Array6.0 chip and scanned. The original data were obtained and then analyzed by CHAS(Chromosome Analysis Suite software. The results were obtained by combining the literature. Results: In 35 cases of congenital heart disease, 6 cases of chromosomal DNA abnormality were detected, the detection rate was 17.14. The results of gene microarray showed that there was 22q11.21 deletion and 22q11.1 duplication in case 1, located between 16055170bp-17023514bp and 18884837bp-21465481bp, respectively. The results of chromosome karyotype analysis showed that there was 22q11.21 deletion in case 3 of 21 trisomy syndrome, which was located between 18793934bp-21431554bp and 22q11 deletion syndrome. In case 4, there were 22q11.21 deletions between 20716876bp-21800471bp and atypical deletions in 22q11. Case 5 was diagnosed as 47% XX. Case 6 had Yp11.31-q11.221 duplication between 2650424bp-16107288bp and Yq11.22q11.23, and between 16167646bp-28799654bp, which was characterized as azoospermic factor deletion. Conclusion: Compared with the traditional methods of cell genetic analysis, the whole genome chip scanning technique can detect chromosomal aberrations at submicroscopic level with high resolution, high throughput and high accuracy, which is beneficial to the screening of pathological copy number variation and pathogenicity genes. It is beneficial to the study of karyotype-phenotypic correlation and can be used as a useful supplement of traditional cell genetic analysis in clinical cell genetic diagnosis.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R725.4

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