新一代测序技术在先天性感音神经性聋中的应用研究  

Application of next generation sequencing in congenital sensorineural deafness

作　　者：徐彬[1] 陈扬[1] 蒋艾[1] 陈操 王凯 郑静 付勇[1] XU Bin CHEN Yang J IANG Ai CHEN Cao WANG Kai ZHENG J ing FU Yong(Department of Otolaryngology, the Children＇s Hospital, Zhejiang University School of Medi- cine, Hangzhou, 310052, China)

机构地区：[1]浙江大学医学院附属儿童医院耳鼻咽喉科,杭州310052

出　　处：《临床耳鼻咽喉头颈外科杂志》2018年第11期811-815,共5页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：国家自然科学基金(面上项目)(No:81470686);浙江省自然科学基金(No:LY14H130001);浙江省医药卫生科学研究基金计划(No:2014KYB077)

摘　　要：目的:应用新一代测序技术(NGS)对门诊确诊为感音神经性聋的患儿进行基因检测,分析耳聋家庭致病基因的携带状况和遗传规律等信息,为遗传咨询、产前诊断、出生缺陷预防的临床实践提供理论基础。方法:收集我科门诊通过病史、听力学检测以及影像学检测确诊为感音神经性聋患儿94例,应用NGS检测与耳聋相关的159个基因外显子区、6个线粒体基因及3个miRNA,再对先证者的父母进行Sanger测序验证,得出先证者及父母耳聋基因表达的情况及相互关系。结果:94例耳聋患儿中,70例重度以上感音神经性聋,13例中重度聋,8例中度聋,3例轻度聋。通过NGS检测出23例携带致聋突变,总突变率为24.5%。GJB2基因突变11例,包括235delC纯合突变6例,235delC和299_300del复合杂合突变4例,235delC和c.176_191del复合杂合突变1例;SLC26A4基因突变5例,其中c.919-2A>G纯合突变2例,c.919-2A>G和c.2168A>G复合杂合突变1例,c.919-2A>G和c.754T>C复合杂合突变1例,c.919-2A>G和c.416-418del复合杂合突变1例;MT-RNR1基因2例;STRC、KCNQ1、USH2A、POU3F4、MITF基因各1例。结论:NGS具有快速、高通量、低成本等特点,可以更好地为临床提供用药、遗传咨询及婚育指导,有效预防和减少遗传性聋的发生。Objective：The next-generation sequencing technology（NGS）was used to perform genetic testing on children diagnosed with sensorineural hearing loss in outpatient clinics.The information on the status and inheritance of disease-causing genes in deafness was analyzed to provide a theoretical basis for genetic counseling,prenatal diagnosis,and birth defects prevention.Method：Ninety-four cases of sensorineural deafness diagnosed by medical history,audiological examination,and imaging examination were collected in our department.Next-generation sequencing was used to detect the region of exons of 159 genes,6 mitochondrial genes,and 3 miRNAs related to deafness.The Sanger sequencing verification was performed on the parents of the probands to find out the gene expression status and relationship between the probands and the parents.Result：Of the 94 children with deafness,70 had severe sensorineural hearing loss,13 had moderate to severe hearing loss,8 had moderate hearing loss,and 3 had mild hearing loss.Twenty-three cases of cariogenic mutations were detected by next generation sequencing,and the total mutation rate was 24.5%.There were 11 cases of GJB2 mutations,including 6 homozygous mutations of 235 delC,4 heterozygous mutations of 235 delC and 299_300 del heterozygous mutation of235 delC and c.176_191 del.There are 5 cases of SLC26 A4 gene mutations,including 2 cases of homozygous mutations of c.919-2 A〉G,one case of c.919-2 A〉G and c.2168 AG compound heterozygous mutations,c.919-2 A〉G and c.754 TC compound heterozygous mutation in 1 case,c.919 2 A〉G and c.416-418 del complex heterozygous mutation in 1 case.There were 2 cases of MT RNR1 mutations,and 1 case of STRC,KCNQ1,USH2 A,POU3 F4,and MITF mutations.Conclusion：The next-generation sequencing has features such as rapid,high throughput,and low cost,which is beneficial to medication guidance,genetic counseling,and marriage and parenting guidance,and help to effectively prevent or reduce the occurrence of hereditary deafness.

关 键 词：新一代测序 听觉丧失 感音神经性 基因 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]