【作者】 赵芳；

【导师】 张芩娜；

【作者基本信息】 山西医科大学 ， 耳鼻咽喉—头颈外科， 2010， 硕士

【摘要】 目的应用遗传性耳聋基因芯片检测母系遗传药物性耳聋家系成员线粒体DNA常见突变类型及频率,筛查聋哑学校学生和门诊耳聋患者中的药物性敏感个体,预测氨基糖苷类抗生素耳毒性损害风险,给予宣教,降低耳聋的发病率。方法收集一母系遗传药物性耳聋家系成员、太原聋哑学校在校学生及门诊散发的非综合征性耳聋患者共计82人的外周静脉血5ml,用酚／氯仿法提取DNA；经PCR扩增和杂交后,用耳聋基因芯片检测国人中常见的药物相关性耳聋基因的2个热点突变,包括线粒体DNA12S rRNA(A1555G,C1494T)。结果在82例样本中,成功提取DNA 81例,其中包括12例家系成员、61例聋哑学校学生及8例门诊患者,共检测出线粒体DNA A1555G突变7例,全部来自药物性耳聋家系,未检测出线粒体DNA C1494T突变阳性。结论线粒体DNA A1555G点突变是导致该家系致聋的主要因素之一,且携带该突变的患者多分布在母系遗传特点的耳聋家系。家系成员的耳聋程度从正常到极重度不等,临床表现为感音性耳聋,遗传方式符合线粒体遗传。更多还原

【Abstract】 Objective Application of the DNA microarray which is able to perform mutation detection of 2 hot-spot mutations in one most common mitochondrial DNA gene simultaneously, including mitochondrial 12S rRNA(A1555G,C1494T),to detect the patients who were the family members of a pedigree with a maternally inherited aminoglycoside antibiotic-induced deafness,deaf students,and clinical patients.Screening and testing for this mutation are effective methods to prevent ototoxicity in A1555G carriers and their maternal family members.Methods Eigty-two non-syndromic hearing loss individuals were included in this study. Blood samples were obtained from 12 family members in a pedigree with a maternally inherited aminoglycoside antibiotic-induced deafness,and 70 sporadic non-syndromic deafness coming from Taiyuan School for the Deaf and our hospital. We collected peripheral blood 5ml and extracted DNA from the isolated leukocytes with phenol/chloroform method,then the mitochondrial DNA target fragments were amplified by polymerase chain reaction (PCR). Their genomic DNA samples were detected with the DNA microarray which is able to perform mutation detection of 2 hot-spot mutations in one most common pathologic mitochondrial DNA gene including mitochondrial 12SrRNA(A1555G,C1494T) simultaneously.Results In the 82 cases of samples,we extracted DNA 81cases successfully,including 12 cases of family members,61 cases of deaf school students and 8 clinical patients. The homoplasmic A1555G mutation was found in 7 individuals of 81 patients,and all the positive patients were coning from the drug-induced deafness pedigree. The C1494Tmutation did not appear in all cases.Conclusion Our results suggested that the mitochondrial DNA 1555G mutation may be one of major factors resulted in aminoglycoside antibiotic-induced deafness in this pedigree, and the 1555G carriers were mainly distributed in the pedigree with maternally inherited deafness.Additionally,this mutation can result in various grades of deafness.The deafness grades ranged from normal to profound. We identified the hereditary model of the pedigree was non-syndromic sensoneural hearing loss mitochondrial inheritance.更多还原