常染色体显性遗传性听神经病家系全外显子组测序分析

发布时间：2018-06-25 01:41

本文选题：听神经病 + 全外显子测序　；参考：《南京医科大学学报(自然科学版)》2017年08期

【摘要】：目的 :在多年围绕1个常染色体显性遗传性非综合征型听神经病家系开展系统分子遗传学研究的基础上,进一步探讨该家系耳聋的致病机制,以期发现新的听神经病致病基因和突变位点。方法:对3例耳聋患者和1例配偶进行全外显子组测序,初步筛选出与家系耳聋相关的候选致病基因。采用PCR-Sanger测序法,检测上述候选基因变异是否与家系表型共分离。最后,以50例与研究家系无关的听力正常人为对照,检测候选致病突变在正常群体中的突变频率和SNPs遗传多态性。结果:全外显子测序分析得到41个候选致病基因突变;用PCR-Sanger测序法对核心家系的9名成员和2名家系外听力正常人进行验证,仅发现1个基因突变(ALOX15B 7942797 CT)与家系耳聋表型共分离。选取50例家系外正常对照的DNA样本对ALOX15B基因进行PCR扩增和序列分析,结果显示有2例听力正常人也检测到该基因的同一变异,提示该变异为SNPs遗传多态性。结论:对核心家系成员的全外显子组测序分析和Sanger测序法验证未发现有意义的突变位点,排除了该家系耳聋由基因编码区突变及Indels致病的可能性。
[Abstract]:Objective: to investigate the pathogenesis of deafness in an autosomal dominant hereditary non-syndromic auditory neuropathy family based on systematic molecular genetic studies for many years. In order to find a new pathogenicity gene and mutation site of auditory neuropathy. Methods: three deafness patients and one spouse were sequenced and the candidate genes associated with deafness were preliminarily screened. PCR-Sanger sequencing was used to detect whether the mutation of the candidate gene was coisolated from the pedigree phenotype. Finally, the mutation frequency and SNPs genetic polymorphism of candidate pathogenetic mutations in normal population were detected in 50 normal subjects unrelated to the study family. Results: 41 candidate gene mutations were obtained by total exon sequencing, 9 members of nuclear family and 2 individuals with normal hearing outside of nuclear family were tested by PCR-Sanger sequencing. Only one gene mutation (ALOX15B 7942797 CT) was coisolated from the deafness phenotype. The ALOX15B gene was amplified by PCR and sequenced from 50 normal controls. The results showed that the same mutation of ALOX15B gene was detected in 2 normal people, indicating that the mutation was SNPs genetic polymorphism. Conclusion: no significant mutation sites were found in the whole exon sequence analysis and Sanger sequencing of nuclear family members, which excluded the possibility of gene coding mutation and Indels pathogenicity of deafness in this family.
【作者单位】：南京医科大学第一附属医院耳鼻咽喉科;南京医科大学生物技术系;
【分类号】：R764

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