肺动脉闭锁遗传学初步研究

发布时间：2018-05-12 04:29

本文选题：肺动脉闭锁 + 染色体　；参考：《华中科技大学》2013年博士论文

【摘要】：第一部分肺动脉闭锁染色体核型分析 [目的]对肺动脉闭锁患儿进行核型检测,分析染色体异常与肺动脉闭锁的关系。 [方法]35例肺动脉闭锁患儿行G显带常染色体检测。 [结果]发现1例合并有21三体综合征,余染色体核型正常。 [结论]21三体综合征可合并有肺动脉闭锁,肺动脉闭锁患儿染色体核型异常并不多见,提示应用常规染色体核型分析无法对这种严重复杂的先心病进行筛查。第二部分肺动脉闭锁TBX1微缺失研究 [目的]对肺动脉闭锁患儿进行TBX1微缺失检测,探讨肺动脉闭锁与TBX1的关系。 [方法]选用荷兰Kreatech公司Poseidon FISH DNA的KBI-40104MD DiGeorge T-boxl(22q11)/22q13(SHANK3),应用染色体原位荧光杂交技术(FISH)对染色体核型正常的34例肺动脉闭锁患儿进行TBX1微缺失检测。 [结果]发现3例TBX1微缺失阳性患儿,阳性率为8.82%。 [结论]TBX1是22q11.2微缺失的核心基因,22q11.2微缺失是肺动脉闭锁的主要致病原因之一,TBX1可能是肺动脉闭锁易感基因。第三部分全外显子测序初步筛选右室流出道梗阻易感基因 [目的]对一个右室流出道梗阻家系中2名成员进行全外显子测序,进行数据分析,初步筛选右室流出道梗阻易感基因。 [方法]收集一个右室流出道梗阻家系中2名成员的静脉血,采用life technology全外显子捕获试剂盒进行捕获,用Life technology solid5500x1高通量测序仪进行高通量测序,测序结果与公共遗传突变数据库(db132SNP)及华中科技大学人类基因组中心正常人外显子数据库进行比对,过滤已报道的常见变异,再将单核苷酸非同义突变进行整合,初步筛选出候选基因。 [结果]结果显示过滤后落在外显子区域的单核苷酸变异(SNP)数目外婆为42727个,孙子为38207个；过滤后落在外显子区域未报道的单核苷酸变异数目外婆为4582个,孙子为4249个；通过ACCESS数据分析发现,有121个新发现单核苷酸变异同时存在于祖孙两人的基因组中。同时使用华中科技大学人类基因组中心已有正常人外显子对照数据库进行筛选,46个变异没有在对照数据库中发现。将46个变异输入到使用ANNOVAR软件预测可能有害的突变位点,将候选基因范围缩小到19个。 [结论]WNK1、KRTAP1-1、UBR2、EPS15、CKAP2、PIGB、DENND4A、 HYDIN、CNTN3、NDUFC2、SAMD9L、KIAA0825、C7orf58、NCKAP5、SPINK5、 FCRL3、TTN、MUC等19个基因可能是右室流出道梗阻易感基因,但须进一步确认。
[Abstract]:Chromosome karyotype analysis of pulmonary atresia Objective: to study the relationship between chromosomal abnormalities and pulmonary atresia in children with pulmonary atresia. [methods] 35 children with pulmonary atresia were detected with G-banded autosomes. [results] A case with trisomy 21 syndrome was found, and the karyotype of cochromosome was normal. [conclusion] trisomy 21 syndrome may be associated with pulmonary atresia, and chromosomal karyotype abnormalities are rare in children with pulmonary atresia, suggesting that routine chromosome karyotype analysis cannot be used to screen this serious complex congenital heart disease. Part II study of TBX1 microdeletion in pulmonary atresia [objective] to investigate the relationship between pulmonary atresia and TBX1 by detecting TBX1 microdeletion in children with pulmonary atresia. [methods] TBX1 microdeletions were detected in 34 children with pulmonary atresia with normal karyotype by using KBI-40104MD DiGeorge T-boxlus 22q11 / 22q13 SHANK3P from Poseidon FISH DNA of Kreatech Company, Netherlands, and fluorescence in situ hybridization (fish). [results] 3 cases of TBX1 microdeletion positive were found, the positive rate was 8.82%. [conclusion] TBX1, the core gene of 22q11.2 microdeletion, is one of the major causes of pulmonary atresia. TBX1 may be a susceptible gene for pulmonary atresia. Part III screening of susceptible genes for right Ventricular outflow tract obstruction by Total Exon sequencing [objective] two members of a family with right ventricular outflow tract obstruction (RVOO) were sequenced, the data were analyzed and the susceptibility genes of RVO were preliminarily screened. [methods] the venous blood of two members of a family with right ventricular outflow tract obstruction was collected and captured by life technology whole exon capture kit. High throughput sequencing was performed by Life technology solid5500x1 high throughput sequencing instrument. The results of sequencing were compared with the common genetic mutation database (Pb132SNPP) and the normal human exon database of the Human Genome Center of Huazhong University of Science and Technology. The reported common mutations were filtered, and the single nucleotide nonsynonymous mutations were integrated. Candidate genes were preliminarily screened. [results] the results showed that the number of single nucleotide mutation (SNPs) in exon region after filtration was 42727 for grandmother and 38207 for grandson, and 4582 for grandmother and 4249 for grandchild after filtering. By ACCESS data analysis, 121 new single nucleotide mutations were found in the genomes of both grandsons and grandsons. At the same time, 46 mutations were not found in the control database using the normal human exon control database in the Human Genome Center of Huazhong University of Science and Technology (Huazhong University of Science and Technology). 46 mutations were inputted to the potentially harmful mutation sites using ANNOVAR software, and the range of candidate genes was reduced to 19. [conclusion] the 19 genes WNK1, KRTAP1-1, UBR2, EPS15, CKAP2PIGBN4A, HYDINC CNTN3NUFC2SAMD9L, KIAA0825, NCKAP5SPINK5, FCRL3TTNMUC, etc., may be susceptible genes to right ventricular outflow tract obstruction, but need to be further confirmed.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R722.11

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