ABCA3基因突变与新生儿呼吸窘迫综合征发病的研究

发布时间：2018-03-18 21:25

本文选题：基因突变　切入点：ABCA3基因　出处：《广西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:研究ATP连接盒转运子A3(ABCA3)基因突变与新生儿呼吸窘迫综合征(RDS)发病的关系,为防治新生儿呼吸窘迫综合征提供基因研究资料。方法:采用病例-对照研究,共纳入广西地区胎龄大于32周的早产儿300例,其中RDS患儿150例(病例组),非RDS患儿150例(对照组)。抽取各研究对象外周静脉血2ml,应用DNA提取试剂盒提取基因组DNA,以每5个DNA建立DNA-pool。采用目标区域捕获技术,以ABCA3为目标基因,通过Agilent液相捕获平台富集目标区域序列,应用Illumina Hiseq2000高通量第二代测序平台对ABCA3基因外显子进行深度重测序并进行数据分析,对错义突变位点用SIFT和Polyphen2软件进行功能预测,确定功能性突变,通过Sanger测序对功能性突变进行基因型验证。应用collapsing统计方法计算总突变的最小等位基因频率(Minor Allele Frequency,MAF)。结果:(1)两组共发现变异位点307种,其中错义变异10种,同义变异15种,内含子变异258种,剪接区域变异2种,上游基因变异位点2种,下游基因变异位点13种,3’端非编码区3种,5’端非编码区4种。(2)病例组未发现突变。对照组发现2种突变:位于Exon 10 c.1009 GA p.V337M和位于Exon 27 c.4149 CG p.I1383M,均为杂合突变。应用Collapsing方法合并突变进行统计:ABCA3基因基因突变在对照组人群的MAF均为0.67%,携带率均为1.33%。结论:ABCA3基因在中国广西地区早产儿人群中突变频率极低,尚无足够依据认为ABCA3基因突变是早产儿RDS发病的高危因素。
[Abstract]:Objective: to study the relationship between the mutation of ATP cassette transporter A3 / ABCA3 gene and the pathogenesis of neonatal respiratory distress syndrome (RDS), and to provide genetic data for the prevention and treatment of neonatal respiratory distress syndrome (RDS). Methods: a case-control study was used to study the relationship between the mutation of ABCA3 gene and the pathogenesis of neonatal respiratory distress syndrome. A total of 300 premature infants with gestational age longer than 32 weeks were included in Guangxi. Among them, 150 cases of RDS (case group) and 150 cases of non RDS (control group) were collected from peripheral venous blood of each study object. Genomic DNAs were extracted by DNA extraction kit, and DNA-pools were established for every 5 DNA. Using target region capture technique, ABCA3 was used as target gene. The target region sequence was enriched by Agilent liquid phase capture platform. The exon of ABCA3 gene was deeply resequenced and analyzed by Illumina Hiseq2000 high-throughput second-generation sequencing platform. The missense mutation sites were predicted by SIFT and Polyphen2 software. Functional mutations were identified, and genotypes of functional mutations were verified by Sanger sequencing. The minimum allelic frequency of total mutations was calculated by collapsing method. The results showed that there were 307 mutation sites in the two groups, of which 10 were missense mutations. There were 15 synonymous mutations, 258 intron variations, 2 splicing region variations, and 2 upstream gene mutation sites. No mutation was found in 13 species of gene mutation loci at the 3'terminal region and 4 species in the 5'terminal noncoding region. In the control group, two mutations were found: Exon 10 c. 1009, GA p.V337M, and Exon 27 c. 4149 CG p.I1383M. all of them were heterozygous mutations. The MAF of the Collapsing gene mutation was 0.67 in the control group, and the carrying rate was 1.33. Conclusion the mutation frequency of the 10% ABCA3 gene is very low in the preterm infants in Guangxi, China. There is no sufficient evidence that ABCA3 gene mutation is a high risk factor for preterm infants with RDS.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R722.1

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