新型多基因检测技术在225例非综合征型耳聋患者基因检测中的应用  

A novel technique for simultaneous multi-gene mutation screening in 225 patients with nonsyndromic hearing loss

作　　者：张迪[1] 段宏[1] 林鹏[2] 程静[3] 王翠翠[1] 马元煦[2] 程岩[2] 赵晖[2] 王巍[2] 徐开旭[2] 韩东一[1] 袁慧军[1,3] 

机构地区：[1]解放军总医院耳鼻咽喉头颈外科耳鼻咽喉科研究所,北京100853 [2]天津市第一中心医院耳鼻咽喉头颈外科天津市耳鼻喉科研究所, 天津300192 [3]第三军医大学西南医院医学遗传中心,重庆400038

出　　处：《中华耳鼻咽喉头颈外科杂志》2016年第3期203-208,共6页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：National Basic Research Program of China (973 Program)，国家重点基础研究发展计划(973计划)项目

摘　　要：目的 利用新型快速多基因检测技术对天津市225例非综合征型耳聋患者进行耳聋基因筛查,在明确分子病因的同时,验证新型多基因检测技术的准确性和有效性.方法 受检人群为来自天津市残联和天津市聋人协会的重度非综合征型耳聋患者,共225例.利用单核苷酸多态分型（single nucleotide polymorphisms scan,SNPscan）技术对GJB2、SLC26A4、线粒体DNA 12S rRNA等三个基因的115个位点进行突变检测.选取部分结果采用Sanger测序加以验证.结果 225例非综合征型耳聋患者中共找到明确基因致聋者111例（49.3％,111/225）.明确GJB2基因突变致聋者56例（24.9％,56/225）,其中纯合突变30例（13.3％,30/225）、复合杂合突变26例（11.6％,26/225）;此外,还发现携带GJB2基因突变的单杂合突变者21例,占受检人群的9.3％ （21/225）.明确SLC26A4突变致聋者50例（22.2％,50/225）,其中纯合突变22例（9.8％,22/225）、复合杂合突变28例（12.4％,28/225）;此外,还发现SLC26A4基因的单杂合突变携带者22例,占受检人群的9.8％ （22/225）.线粒体DNA 12S rRNA A1555G突变5例（2.2％,5/225）,未发现1494C＞T突变.测序结果经Sanger测序验证,符合率达100％.225例患者的样本从提取DNA到得到测序结果共耗时1个工作日（8 h）,每个样本花费人民币160元左右.结论 SNPscan耳聋基因诊断技术可以对耳聋患者进行准确、快速和经济有效的分子病因筛查,有望成为大规模遗传性耳聋基因检测的有效手段.Objective Using simultaneous muhi-gene mutation screening to investigate the new method molecular epidemiological basis of 225 patients with nonsyndromic hearing loss in Tianjin, and verifying the for simultaneous multi-gene mutation screening.Methods Two hundred and twenty-five patients with severe non-syndromic deafness from Tianjin CDPF and Association of the Deaf were included in the study.The single nucleotide polymorphisms scan, （SNPscan）technique was used for screening the 115 spots mutations in three common deafness-related genes（GJB2, SLC26A4, mtDNA 12S rRNA） of patients with nonsyndromic hearing loss in Tianjin.We verified the results by Sanger sequencing.Results Among the 225 patients,there were 111 cases of deafness caused by mutation （49.3%）.Using this method, up to 50% of the patients in our study were identified to have hereditary HL caused by mutations in the three genes.56 patients with the GJB2 mutations were detected （24.9%）, including 30 cases of homozygous mutations （13.3%）, 26 patients （11.6%） of compound heterozygous mutations, and 21 cases （9.33%） of single heterozygous mutations.50 patients with the SLC26A4 mutations were detected（22.2%）, including 22 cases of homozygous mutations（9.8%）, 28 patients （12.4%） of compound heterozygous mutations,and 22 cases （9.8%） of single heterozygous mutations.mtDNA 12S rRNA A1555G mutation was detected in 5 patients （2.2%）.mtDNA 12S rRNA 1494C 〉 T mutation was not detected.We verified the results by Sanger sequencing.The accuracy of the sequencing results was 100%.The SNPscan cost eight hours and 160 yuan （each sample）.Conclusions Applying SNPscan technology can be accurate, rapid and cost-effective diagnostic screening in patients with hearing loss for etiology investigation.It is expected to become an effective means of large-scale genetic testing for hereditary deafness.

关 键 词：听觉丧失 基因测定 

分 类 号：R764.43[医药卫生—耳鼻咽喉科] R440[医药卫生—临床医学]