摘要
目的:对遗传性非综合征型耳聋患者进行中国人常见的耳聋相关基因的突变分析,以明确其分子病因。方法:门诊收集非综合征耳聋患者22名的外周血样本,常规方法提取基因组DNA,PCR扩增GJB2基因和线粒体12SrRNA基因全序列,以及SLC26A4基因7+8外显子和19外显子序列。所有PCR扩增产物经DNA测序分析,测序结果提交NCBIGenBank数据库进行比对,从而对耳聋相关基因的突变进行分析。结果:22名非综合征耳聋患者中共检测到3种GJB2基因的突变:235delC纯合突变2例;235delC杂合突变3例及176del16bp和299delAT复合突变1例;mtDNA12SrRNA基因检出有2种已知致病突变:A1555G2例和C1494T2例;SLC26A4基因有IVS7-2A>G杂合突变2例,纯合突变1例。结论:PCR扩增结合DNA测序是耳聋基因诊断的经典和有效的方法,能确诊非综合征型耳聋患者的分子病因,为临床诊断和治疗提供依据。但该方法存在不能定性、耗时费力、所需成本昂贵且不能同时对不同基因的多个突变位点进行检测等缺点。因此,临床的耳聋基因诊断,需要操作简便、结果准确、通量高、价格低廉的新手段。
Objective：To investigate the mutation of non-syndromic sensorineural hearing loss （NSHL） and identify its molecular etiopathogenisis. Methods：Peripheral blood samples were obtained from 22 NSHL patients collected by out-patient clinic. Their genomic DNA were extracted from peripheral blood by extraction kits to undergo polymerase chain reaction and sequencing so as to detect the mutations of GJB2,SLC26A4 and mitochondrial 12SrRNA gene. Results：Of 22 patients,three mutations of GJB2 gene were found：2 have GJB2 235delC heterozygous mutation,2 have 235delC homozygous mutation and 1 have 176del16bp＋299delAT double mutation;2 mutaions was found out in MtDNA 12SrRNA gene：C1494T and A1555G;SLC26A4 gene have 1 IVS7-2 homozygous and 2 heterozygous mutations. Conclusion：The PCR combined DNA sequencing is a classic and effective method in genetic diagnostic of hereditary deafness,able to identify the molecular etiopathogenisis of NSHL. But this method exist many shortcomings,such as unable to qualitate, time and money consuming. Furthermore,it can not detect multiple mutations of different genes at the same time. Therefore,a new method,which has advantages in genetic diagnosis of NSHL,such as low time and money consuming,high performance and accuracy, must to be developed. These advantages make it fit to be used in clinic gene testing of hereditary deafness.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第3期390-393,共4页
Journal of Nanjing Medical University(Natural Sciences)
基金
江苏省高等学校大学生实践创新训练计划2008年立项资助项目(No.267)
南京医科大学校基金(09NJ-MUM005)资助
