毛球线粒体DNA的检测及临床应用研究  

The clinical evaluation for the mitochondrial DNA detected from hair follicles

作　　者：李霞[1] 周新[1] 李杰[1] 郭清莲[1] 鄢盛恺[2] 鲁敏翔[1] 

机构地区：[1]武汉大学中南医院基因诊断中心,430071 [2]中国医学科学院北京协和医院检验科

出　　处：《中华检验医学杂志》2005年第6期595-598,共4页Chinese Journal of Laboratory Medicine

基　　金：湖北省自然科学基金资助项目(2004ABA163);武汉市科委重点资助项目(20016009107)

摘　　要：目的比较肌肉、血液和毛球线粒体DNA(mtDNA)的异质性多态水平,探讨毛球在线粒体相关疾病基因分析中的应用价值。方法收集50名无血缘关系汉族个体的肌肉、血液和毛发标本,采用聚合酶链反应(PCR)扩增mtDNA的高变区Ⅰ(HVⅠ)1603316368bp片段并测序,比较3种不同组织的异质性多态水平。同时采用PCR限制性长度多态性(RFLP)检测3例线粒体脑肌病合并乳酸血症及卒中发作综合征(MELAS)患者的mtDNAtRNAleu基因nt.3243(A→G)突变,比较不同组织的突变检测率。结果肌肉标本异质性检测率为18%(9/50),毛球为16%(8/50),两者较接近,但均明显高于血液的10%检测率(5/50)。3例MELAS患者的肌肉和毛球标本均检出A3243G突变,仅两例患者的血液标本中检出此突变。结论毛球mtDNA的异质性多态水平接近肌肉,高于血液;毛球标本的线粒体点突变检测率高于血液,因此在线粒体相关疾病的基因分析中具有较高的临床应用价值。Objective To compare the levels of heteroplasmic polymorphism of mitochondrial DNA (mtDNA) in muscle, blood and hair follicles and to study the clinical application value of hair follicles for the genetic analysis of mitochondrial diseases.Methods Muscle, blood and hair follicles were collected from 50 unrelated Han population. The fragment of 16033-16368bp in mtDNA hypervariable regionsⅠ(HVⅠ) was amplified using PCR and sequenced subsequently. Then, the levels of heteroplasmic polymorphism in these three different tissues were compared. Using PCR-RFLP, the nt.3243 (A→G) point mutation in mtDNA tRNA~ leu gene of three mitochondrial encephalomypathy, lactic acidosis and atrokelike episodes (MELAS) patients was also tested, and the mutation detected rate of these three tissues was analyzed.Results The detection rate of heteroplasmic polymorphism in muscle was 18%(9/50) which was close to 16%(8/50) in hair follicles. But both of them were higher than in blood 10%(5/50). The A3243G point mutation was detected in muscle and hair follicles in all MELAS patients. In only two of the three patients was the mutation detected in blood samples.Conclusion The level of mtDNA heteroplasmic polymorphism in hair follicles was close to that in muscle and higher than in blood. The point mutation detection rate of hair follicles was higher than that of blood. So, hair follicles have the potent clinical application value for the genetic analysis of mitochondrial diseases.

关 键 词：临床应用研究 线粒体DNA 聚合酶链反应(PCR) 限制性长度多态性 mtDNA 线粒体脑肌病 临床应用价值 基因分析 相关疾病 毛球标本 无血缘关系 MELAS 异质性 检测率 高变区Ⅰ 乳酸血症 血液标本 肌肉 水平 患者 综合征 G突变 

分 类 号：R446.9[医药卫生—诊断学]