运用耳聋基因隐性突变携带者重测序策略纠正假阳性变异的致病性误判

发布时间：2018-03-17 16:10

本文选题：外显子测序　切入点：耳聋　出处：《上海交通大学学报(医学版)》2017年11期 　论文类型：期刊论文

【摘要】：目的·运用隐性突变携带者重测序策略发现并归类耳聋致病基因罕见良性变异,为遗传性耳聋的高通量二代测序结果解读提供良性多态参考数据库。方法·在已明确隐性耳聋基因双等位基因致病突变的耳聋患者的正常听力亲属中进行相应突变筛查,发现单杂合致病突变的携带者。运用靶向捕获二代测序技术在这些携带者中对相应耳聋基因进行外显子重测序,对所发现的对侧等位基因变异进行非致病性推断。结果·共30位正常听力的耳聋病例亲属被明确为某一已知耳聋基因隐性致病突变的杂合携带者。通过相应基因的靶向二代测序,在对侧等位基因上共发现32个非同义变异,在隐性完全外显的遗传模式下可被归类为良性非致病变异。其中SLC26A4基因p.A434T变异、LOXHD1基因p.R266Q变异、MYO15A基因p.K96Q变异、GJB2基因p.T123N变异及CDH23基因p.V1299I变异,共5个罕见变异等位基因频率小于0.005,部分变异被Polyphen-2、PROVEAN、SIFT或Mutation Taster软件预测为致病突变,或被耳聋变异数据库或人类基因组突变数据库记载为致病突变。结论·运用隐性突变携带者重测序策略可有效纠正部分被误判为致病的罕见良性变异,提高二代测序在单基因隐性遗传病病因诊断中的准确性和有效性。
[Abstract]:Objective: to identify and classify rare benign mutation of deafness gene by recessive mutation carrier resequencing strategy. To provide a benign polymorphic reference database for the interpretation of high-throughput second-generation sequencing results of hereditary deafness. Methods: to screen the mutation in normal hearing relatives of deafness patients who had identified the double allelic mutations of recessive deafness gene. We found the carriers of single heterozygous mutations. The corresponding deafness genes were sequenced in these carriers by targeting capture second-generation sequencing technique. Results: 30 relatives of normal hearing loss patients were identified as heterozygous carriers of a known recessive mutation of deafness gene. Of the second generation, A total of 32 non-synonymous variants were found in the contralateral alleles. The SLC26A4 gene p.A434T mutation and the LOXHD1 gene p.R266Q mutation, the MYO15A gene p.K96Q mutation, the GJB2 gene p.T123N mutation and the CDH23 gene p.V1299I mutation can be classified as benign non-pathogenic variation. The frequencies of 5 rare alleles were less than 0.005. Some of the mutations were predicted to be pathogenic mutations by Polyphen-2ProVEAN sift or Mutation Taster software. Conclusion: recessive mutation carrier resequencing strategy can effectively correct some of the rare benign mutations that were misjudged as pathogenicity by deafness mutation database or human genome mutation database. To improve the accuracy and effectiveness of second generation sequencing in etiological diagnosis of single gene recessive hereditary disease.
【作者单位】：上海交通大学医学院附属第九人民医院耳鼻咽喉-头颈外科上海交通大学医学院耳科学研究所上海市耳鼻疾病转化医学重点实验室;
【基金】：上海市教育委员会高峰高原学科建设计划(20152519)~~
【分类号】：R764.43

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