十五项耳聋基因芯片联合一代测序检测聋儿家庭的致聋变异分析  

DNA Microarray and Sanger Sequencing in Genetic Diagnosis of Nonsyndromic Hearing Impairment Families

作　　者：林颖[1] 周枫[1] 黄利芬[1] 张群慧[1] 王兴君[1] 王淑杰 于锋[1] LIN Ying;ZHOU Feng;HUANG Lifen;ZHANG Qunhui;WANG Xingjun;WANG Shujie;YU Feng*(Department of Otolaryngology Head and Neck Surgery,Guangzhou Otolaryngology Head and Neck Surgery Hospital,Guangzhou Medical University,Guangzhou,510620,China)

机构地区：[1]广州市第十二人民医院(广州医科大学附属广州市耳鼻咽喉头颈外科医院)

出　　处：《中华耳科学杂志》2019年第5期716-721,共6页Chinese Journal of Otology

基　　金：广州市科技计划项目（201704020178）;广州市卫生和计划生育科技项目（2019A010040）;广州市社会组织公益创投项目（E59)~~

摘　　要：目的应用十五项耳聋基因芯片联合一代测序技术对非综合征型耳聋家庭易感基因进行检测,揭示常见的耳聋分子病因构成,并验证基因芯片技术的准确性及有效性。方法对广州市两个特殊听障机构的80组聋儿家庭(共241例)进行病史采集、听力评估,应用十五项耳聋基因芯片技术进行GJB2(c.35delG,c.176del16bp,c.235delC及c.299_300delAT),GJB3(c.538C>T),SLC26A4(c.919-2A>G,c.2168A>G,c.1229C>T,c.1975G>C,c.1174A>T,c.1226G>A,c.2027T>A和c.IVS15+5G>A)和MT-RNR1(m.1555A>G,m.1494C>T)的检测,并用Sanger法对变异阳性家庭进行序列测定。结果在80组聋儿家庭中,基因芯片检测出GJB2及SLC26A4基因变异共62例,总检出率为25.73%(62/241),GJB2 c.235delC(34/62)与SLC26A4 c.919-2A>G(16/62)热点变异居多。Sanger法测序结果显示:携带GJB2及SLC26A4基因共73例,总检出率为30.30%(73/241),其中GJB2 49例,SLC26A4 24例;基因芯片检测出的基因变异经一代测序验证,符合率达100%;除基因芯片包含的位点外,还检测出部分GJB2 c.109G>A、c.79G>A、c.341A>G变异,及SLC26A4 c.259G>T、c.754T>C变异。结论十五项耳聋基因芯片联合一代测序技术可以提高非综合征型耳聋的变异检出率,此基因芯片准确率高,适合快速、大规模检测;广州市此特殊听障机构的耳聋相关基因热点变异仍以GJB2 c.235delC与SLC26A4 c.919-2A>G最常见。Objective To detect susceptible genes in nonsyndromic hearing impairment families using DNA microarray kit targeting 15 deafness mutations in combination with sanger sequencing,and verify accuracy and validity of the technologies.Method A total of 80 families(241 individuals)from two hearing impairment institutions in Guangzhou received detailed medical history,hearing test and DNA microarray testing.Individuals showing mutations underwent Sanger sequencing.Result Among the 80 families,DNA microarray showed mutations involving the GJB2 and SLC26A4 genes in 25 families(62 individuals),with the hot loci GJB2 c.235delC and SLC26A4 c.919-2A>G in the majority of cases.Sanger sequencing showed mutations in 30.30%(73/241)of the subjects,including 49 cases involving the GJB2 and 24 cases involving the SLC26A4 genes.The results of DNA microarray were 100%consistent with those of Sanger Sequencing.In addition to loci contained in the microarray,Sanger sequencing also revealed SNP of GJB2(including c.79G>A,c.341A>G and c.109G>A)and mutations of SLC26A4(including c.259G>T and c.754T>C).Conclusion Because of its high accuracy and fast speed,the DNA microarray kit for 15 target deafness mutations can be used in large scale screenings,and can improve the detection of deafness gene mutations when combined with Sanger Sequencing.GJB2 c.235delC and SLC26A4 c.919-2A>G remain the common causative mutation loci among these patients from the two hearing impairment institutions of Guangzhou.

关 键 词：基因芯片 Sanger法 序列测定 非综合征型耳聋 易感基因 

分 类 号：R764[医药卫生—耳鼻咽喉科]