新一代测序技术在先天性感音神经性聋中的应用研究

徐彬; 陈扬; 蒋艾; 陈操; 王凯; 郑静; 付勇

doi:10.13201/j.issn.1001-1781.2018.11.003

新一代测序技术在先天性感音神经性聋中的应用研究

- 浙江大学医学院附属儿童医院耳鼻咽喉科(杭州, 310052)
基金项目:
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）

详细信息

通讯作者: 付勇,E-mail:1307022@zju.edu.cn

中图分类号: R764.43

收稿日期: 2018-04-12

Application of next generation sequencing in congenital sensorineural deafness

- Department of Otolaryngology, the Children's Hospital, Zhejiang University School of Medicine, Hangzhou, 310052, China

More Information

Corresponding author: FU Yong, E-mail: 1307022@zju.edu.cn

Received Date: 12 April 2018

摘要

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。
- 新一代测序 /
- 听觉丧失,感音神经性 /
- 基因
Abstract: 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 235delC, 4 heterozygous mutations of 235delC and 299_300del heterozygous mutation of 235delC and c.176_191del. There are 5 cases of SLC26A4 gene mutations, including 2 cases of homozygous mutations of c.919-2A>G, one case of c.919-2A>G and c.2168A>G compound heterozygous mutations, c.919-2A>G and c.754T>C compound heterozygous mutation in 1 case, c.919 2A>G and c.416-418del complex heterozygous mutation in 1 case. There were 2 cases of MT RNR1 mutations, and 1 case of STRC, KCNQ1, USH2A, POU3F4, 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.
- next generation sequencing /
- hearing loss, sensorineural /
- gene

HTML全文

参考文献(19)

[1]	HUANG X Z, WANG J B, KONG W J, et al.Practice of Otorhinolaryngology-Head and Neck Surgery[M].2nd.Beijing:People's Medical Publishing House, 2010:976-976.
[2]	ACMG.Genetics evaluation guidelines for the etiologic diagnosis ofcongenital hearing loss.Genetic Evaluation of Congenital Hearing Loss Expert Panel.ACMG statement[J].Genet Med, 2002, 4:162-171.
[3]	戴朴, 袁永一, 辛凤.耳聋基因诊断与遗传咨询[M].北京:人民卫生出版社, 2017:208-209.
[4]	戴朴.遗传性耳聋的预防和阻断[J].中华医学杂志, 2007, 87(40):2811-2813.
[5]	DAI Z Y, SUN B C, HUANG S S, et al.Correlation analysis of phenotype and genotype of GJB2 in patients with non-syndromic hearing loss in China[J].Gene, 2015, 570:272-226.
[6]	GRILLO A P, DE OLIVEIRA F M, DE CARVALHO G Q.Single nucleotide polymorphisms of the GJB2 and GJB6 genes are associated with autosomalrecessive nonsyndromic hearing loss[J].Biomed Res Int, 2015, 2015:318727.
[7]	SHI L, CHEN J.Prevalence of GJB2 gene mutation in330cochlear implant patients in the Jiangsu province[J].J Laryngol Otol, 2016, 130:902-906.
[8]	WOLF A, FROHNE A, ALLEN M, et al.A Novel Mutation in SLC26A4 Causes Nonsyndromic Autosomal Recessive Hearing Impairment[J].Otol Neurotol, 2017, 38:173-179.
[9]	JUNG J, SEO Y W, CHOI J Y, et al.Vestibular function is associated with residual low-frequency hearing loss in patients with bi-allelic mutations in the SLC26A4 gene[J].Hear Res, 2016, 335:33-39.
[10]	ROSE J, MUSKETT J A, KING K A, et al.Hearing loss associated with enlarged vestibular aqueduct and zero or one mutant allele of SLC26A4[J].Laryngoscope, 2017, 127:E238-E243.
[11]	YUAN Y, GUO W, TANG J, et al.Molecular epidemiology and functional assessment of novel allelic variants of SLC26A4 in non-syndromic hearing loss patients with enlarged vestibular aqueduct in China[J].PLoS One, 2012, 7:e49984.
[12]	MÉNDEZ-VIDAL C, GONZÁLEZ-DEL POZO M, VELA-BOZA A, et al.Whole-exome sequencing identifies novel compound heterozygous mutations in USH2A in Spanish patients with autosomal recessive retinitis pigmentosa[J].Mol Vis, 2013, 19:2187-2195.
[13]	MAERKER T, VAN WIJK E, OVERLACK N, et al.A novel Usher protein network at the periciliary reloading point between molecular transport machineries in vertebrate photoreceptor cells[J].Hum Mol Genet, 2008, 17:71-86.
[14]	EUDY J D, WESTON M D, YAO S, et al.Mutation of a gene encoding aprotein with extracellular m atrix motifs in Usher syndrome type IIa[J].Science, 1998, 280:1753-1757.
[15]	VYAS B, PURI R D, NAMBOODIRI N, et al.KCNQ1mutations associated with Jervell and lange-nielsen syndrome and autosomal recessive Romano-Ward syndrome in India-expanding the spectrum of long QT syndrome type 1[J].Am J Med Genet A, 2016, 170:1510-1519.
[16]	DUPONT M, JONES EM, XU M, et al.Investigating the disease association of USH2Ap.C759 Fvariant by leveraging large retinitispigmentosa cohort data[J].Ophthalmic Genet, 2018, 39:291-292.
[17]	NAKASHIMA K, KUSAKAWA I, DUFLOU J, et al.A left ventricular noncompaction in a patient with long QT syndrome caused by a KCNQ1 mutation:a case report[J].Heart Vessels, 2013, 28:126-129.
[18]	ARGENIO V, FRISSO G, PRECONE V, et al.DNA sequence capture and next-generation sequencing for the molecular diagnosis of genetic cardiomypathies[J].J Mol Diagn, 2014, 16:32-44.
[19]	PINGAULT V, ENTE D, DASTOT-LE MOAL F, et al.Review and update ofmutations causing Waardenburg syndrome[J].Hum Mutat, 2010, 31:391-406.