口腔拭子在药物性耳聋基因检测中的应用  

Applications of genes extracted from buccal swab for screening mitochondrial DNA A1555G and C1494T mutation

作　　者：张琼敏[1] 郑静[2] 宋攀攀[1] 朱翌[1] 陈晓云[1] 管敏鑫[2] 

机构地区：[1]温州医学院附属第一医院耳鼻咽喉科,浙江温州325000 [2]温州医学院生命科学学院,浙江温州325035

出　　处：《温州医学院学报》2011年第5期409-412,共4页Journal of Wenzhou Medical College

基　　金：国家"973"重大基础研究前期研究专项(2004CCA02200);国家自然科学基金资助项目(81070794);浙江省医药卫生科学研究基金资助项目(2006A100);浙江省"钱江人才计划"择优资助项目(2006R10021);浙江省重大科技专项社会发展项目(2007C13021)

摘　　要：目的:探讨口腔黏膜上皮细胞基因组DNA在药物性耳聋基因A1555G及C1494T突变检测中的应用,寻求基因检测中更简单快捷并无损的样本来源。方法:采集97例来自温州特殊教育学校非综合征型耳聋学生的口腔拭子和外周血,分别提取基因组DNA,进行浓度与纯度的测定、基因扩增及产物测序,比较2种样本来源及2种方法提取的基因组DNA的浓度、纯度、扩增成功率,并将测序结果与人类线粒体DNA剑桥参考序列进行比对。结果:口腔拭子DNA的手工法提取浓度(42.5±41.7)ng/mL与试剂盒法DNA浓度(44.8±43.4)ng/mL差异无统计学意义(P>0.05);手工法DNA纯度(1.45±0.73)低于试剂盒法(1.87±0.87),两组间差异有统计学意义(P<0.05);二者扩增成功率与外周血差异均无统计学意义(P>0.05);口腔拭子DNA的药物性耳聋基因扩增产物长短一致,测序结果完全相符,均显示A1555G突变阳性2例,阴性95例;C1494T突变97例均阴性。结论:无损性样本口腔拭子扩增成功率高,诊断结果可靠,可替代外周血作为药物性耳聋基因检测的DNA来源,具有一定的临床应用价值。Objective： To study the applications of genes extracted from buccal swab for screening mitochondrial DNA A1555G and C1494T mutation,and search for a simple and non-invasive kind of gene source.Methods： Determine the purity and concentration of genes from buccal swabs via Manual sampling and Reagent kits and compare them with that of peripheral blood from 97 nonsyndromic hearing impairment students at Wenzhou Special Education School.Amplify and sequence the DNA from the two different kinds of samples and then compare the results in NCBI.Results： The concentration of the genes from buccal swabs via Manual sampling（42.5±41.7）ng/L had no statistically difference from that via Reagent kits（44.8±43.4）ng/L（P 0.05）.The purity of the former（1.45±0.73）was lower than that of the later（1.87±0.87）（P 0.05）.The amplification success rate of both had no statistically difference from that of periph-eral blood（P 0.05）.The amplification of buccal swab DNA and blood have nearly same length and the results profit to the standard sequence banded in NCBI,2 cases were A1555G positive and 95 cases were A1555G negative.all were C1494T negative.Conclusion： The amplification success rate of buccal swabs is equal to the peripheral blood and the result is credible.Buccal swabs as a source of genes is of higher clinical value,which can be a stubstitution of blood.

关 键 词：无损性 口腔黏膜拭子 药物性耳聋 基因检测 基因扩增 

分 类 号：R394.33[医药卫生—医学遗传学]