中国人一先天性巨结肠家系全外显子测序分析

发布时间：2018-05-17 08:31

本文选题：先天性巨结肠 + 单核苷酸　；参考：《华中科技大学》2012年博士论文

【摘要】：目的采用新一代测序技术对一个汉族巨结肠家系中2例患者进行全外显子测序，测序数据进行多步骤过滤分析，初步筛选先天性巨结肠的易感基因。方法收集一个先天性巨结肠家系中母子2个患者的静脉血采用全外显子捕获和新一代测序技术进行高通量测序，测序结果与人类HAPMAP8，dbSNP130和1000Genome Project数据库进行比对过滤已报道的常见变异，再过滤掉同义突变，将母子单核苷酸非同义突变进行整合，并通过Sanger法测序排除外显子测序的假阳性结果，初步筛选出候选基因。结果测序结果通过原始质量评分过滤，母子两个患者共得到8.4G的数据。过滤掉不在外显子区域的单核苷酸变化(SNVs)，落在外显子区域的单核苷酸变化数目母亲为13948个，，儿子为13856个；过滤掉公共数据库（HAPMAP8，dbSNP130和1000Genome Project）的常见变异，落在外显子区域的未报道的单核苷酸变化数目母亲为3472个，儿子为3345个；过滤掉同义突变，非同义突变的单核苷酸变化数目母亲为470个，儿子为458个；整合母子两个共有的非同义突变数目筛选出17个基因。Sanger法排除外显子测序C22orf42(G＞A)假阳性结果，最后得到16个候选基因。结论NRG3；LAMA3；BRIP1；JARID2； KRT6A；LEPREL1；OR8J3；PLA2G4C；PRBP4；RNF10；SPRY1；TMPRSS11E；VARS2；NBPF16；GSTM4；PRSS116个基因可能为巨结肠的易感基因，但须进一步确认。
[Abstract]:Objective to detect the susceptibility genes of Hirschsprung's colon (Hirschsprung's disease) by sequencing two patients with Hirschsprung's disease (Hirschsprung's disease) in a Han Chinese pedigree by multistep filtration analysis. Methods the venous blood of two patients with Hirschsprung's disease was collected from a family of Hirschsprung's disease and high throughput sequencing was performed by whole exon capture and a new generation of sequencing techniques. The sequencing results were compared with human HAPMAP8dbSNP130 and 1000Genome Project database to filter the reported common mutations, then filter out synonymous mutations, integrate mother and son single nucleotide nonsynonymous mutations, and exclude the false positive results of exon sequencing by Sanger sequencing. Candidate genes were preliminarily screened. Results the sequencing results were filtered by the original quality score, and the data of the two patients were 8.4G. The number of single nucleotide variations in the exon region was 13948 for mothers and 13,856 for sons, and the common variations in the common database, HAPMAP8dbSNP130 and 1000Genome projects, were filtered out. The number of unreported single nucleotide changes in the exon region was 3472 for mother and 3345 for son, 470 for mother and 458 for son, filtering out synonymous mutation and non-synonymous mutation. 17 genes were screened by Sanger method to exclude exon sequencing C22orf42(G > A) false positive results were obtained. Finally, 16 candidate genes were obtained. Conclusion NRG3, Lama 3, BRIP1, JARID2, KRT6A, LEPREL1, OR8J3G4G4, PRBP4, RNF10, SPRY1, TMPRSS11EVARS2, NBPF16GSTM4PRSS116, may be the susceptibility genes of megacolon, 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】：2012
【分类号】：R722.11

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