17000名新生儿遗传性耳聋基因突变筛查  

Screening of common deafness gene mutations in 17 000 Chinese newborns from Chengdu based on microarray analysis

作　　者：吕康模 熊业华 俞皓 邹玲 冉隆荣 刘德顺 殷勤 徐应文 方雪 宋祖玲 黄丽佳[4] 谭大勇 张治位 

机构地区：[1]成都市妇女儿童中心医院产前诊断中心,610091 [2]成都博奥独立医学实验室有限公司 [3]成都市双流县妇幼保健院 [4]成都市龙泉驿区妇幼保健院 [5]成都市温江区妇幼保健院

出　　处：《中华医学遗传学杂志》2014年第5期547-552,共6页Chinese Journal of Medical Genetics

摘　　要：目的应用遗传性耳聋基因芯片对新生儿进行致聋基因突变筛查，评估成都地区新生儿中常见遗传性耳聋基因的突变频率和类型。方法收集成都地区2012年8月至2013年6月送检的17000名新生儿足跟血制成干血斑，应用微阵列芯片法检测中国人常见的4个致聋基因的9个突变位点，包括GJB2基因（35delG、176dell6、235delC、299delAT）、GJB3基因（538C〉T）、SLC26A4基因（IVS7—2A〉G、2168A〉G）、线粒体DNA12SrRNA基因（1555A〉G、1494c〉T）。对所有阳性结果和随机抽取2％的阴性结果进行测序验证。结果17000名新生儿中检出突变者542例，其中GrJB2235delC254例、176del1615例、299delAT55例，GJB3538C〉T23例，SLC26A42168A〉G17例、IVS7—2A〉G128例，线粒体12SrRNA1494C〉T4例、1555A〉G42例（含2例复合性突变中有1555A〉G均质突变）；另外检出复合性突变6例，分别为GJB2235delC杂合突变伴有SLC26A4IVS7—2杂合突变和线粒体DNA12SrRNA1555A〉G均质突变各1例，GJB2299delAT杂合突变伴有GJB3538C〉T杂合突变和线粒体DNA12SrRNA1555A〉G均质突变各1例、伴SLCg6A4IVS7—2A〉G杂合突变2例。测序结果与芯片结果一致。在受检的新生儿中，耳聋基因GJB2，GJB3，SLC26A4和12SrRNA突变总携带率达到3．19％；致药物性耳聋的线粒体DNA基因突变率达到0．27％；GJB2基因与SLCg6A4基因突变携带率达2．76％，占基因突变携带人群的86．5％。结论在无耳聋家族史的新生儿中，4个致聋基因突变均有携带者，GJB2基因突变高于SLC26A4基因突变，两者占比最大。致药物性耳聋的线粒体DNA12SrRNA基因突变在新生儿用药前及时发现，并终生避免接触氨基糖苷类药物，将有利于防止“一针致聋”的发生。在新生儿中开展耳聋基因筛查，对遗传性耳聋的早期诊断和预防，以及成年后的婚育指导具有重要意义。Objective To achieve early diagnosis for inheritable hearing loss and determine carrier rate of deafness causing gene mutations in order to provide information for premarital, prenatal and postnatal genetic counseling. Methods A total of 17 000 dried heel blood spots of normal newborns in Chengdu were collected with informed consent obtained from their parents. Genomic DNA was extracted from dried blood spots using Qiagen DNA extraction kits. Microarrays with 9 common mutation loci of 4 deafness-associated genes in Chinese population were used. Nine hot mutations including GJB2 （35delG, 176de116, 235delC and 299delAT）, GJB3 （538C 〉 T）, SLC26A4 （IVS 7 - 2 A 〉 G, 2168 A 〉 G）, and mitochondrial DNA 12S rRNA（1555 A〉G, 1494 C〉T） were detected by PCR amplification and microarray hybridization. Mutations detected by microarray were verified by Sanger DNA sequencing. Results Of the 17 000 new-borns, 542 neonates had mutations of the 4 genes. Heterozygous mutations of GJB2, at 235delC, 299delAT, and 176de116 were identified in 254, 55, and 15 newborns, respectively. Two newborns had homozygous mutation of GJB2, 235delC. Heterozygous mutations at 538C〉T of GJB3, 2168A〉G and IVS 7--2A〉G of SLC26A4 were found in 23, 17 and 128 newborns, respectively. For mutation analysis of mitochondrial DNA 12S rRNA, 1494C〉 T and 1555A 〉 G were homogeneous mutations in 4 and 42 neonates, respectively. In addition, 6 complexity mutations were detected, which demonstrated that one newborn had heterozygous mutations at GJB2 235delC and SLC26A4, IVS7- 2A〉G, one had heterozygous mutation GJB2 235delC and 12S rRNA homogeneous mutation, 1555 A^G, one heterozygous mutations at GJB2, 299delAT, and GJB3, 538C〉T, one at GJB2, 299delAT and 12S rRNA, 1555 A〉 G, two at GJB2, 299delAT, and SLC26A4, IVST--2A〉G. All mutations as above were confirmed by DNA sequencing. Conclusion The total mutation carrier rate of the 4 deafness genes is 3.19% in healthy newborns at Chengdu. Mutations of GJB2 and SLAC26A4 are majo

关 键 词：遗传性耳聋 GJB2基因 SLC26A4基因 12SRRNA基因 基因突变 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]