摘要
目的探讨线粒体脑肌病的分子生物学诊断方法。方法以2对寡核苷酸为引物对3例患者血液和骨骼肌DNA进行聚合酶链反应(PCR)扩增,在PCR反应最后一个循环加入-α32P-dCTP,产物经Apa和BglⅠ酶酶切后进行聚丙烯酰胺凝胶电泳,放射自显影,吸光度扫描仪扫描分析。结果病例1和2线粒体DNA存在A3243G点突变,病例1血液和骨骼肌突变型mtDNA分别为34.2%和66.8%,病例2为37.5%和73.3%;病例3存在A8344G点突变,血液和骨骼肌突变型mtDNA分别为46.2%和76.4%。结论PCR结合限制性内切酶分析可作为线粒体脑肌病的分子诊断方法,可为其确诊提供有力的分子遗传学证据。
Objective To investigate molecular diagnosis method for mitochondrial encephalomyopathy. Methods The mtDNA mutations were detected by polymerase chain reaction （PCR） assay coupled to restriction digestion. The radioactivity （α-32P-dCTP） was added at the last cycle of PCR, The products ran on a 8% polyacrylamide gel after restriction digestion with Apa Ⅰ and Bgl Ⅰ,respectively,and then the percentage of mutant mtDNA was detected by antoradiography, Results MtDNA A3243G mutation was identified in tissue of patients 1 and 2, and their percentages of mtDNA A3243G were 66.8 % and 73.3 % in muscle as well as 34.2 % and 37.5 % in blood, respectively. The A8344G mutation was heteroplasmic in patient 3, and the.percentages of mtDNA A8344G in muscle and blood were 46.2 % and 76.4 %, respectively. Conclusion PCR-restriction enzyme digestion can act as the method for molecular diagnosis of mitochondrial encephalomyopathy.
出处
《山西医药杂志》
CAS
2006年第5期393-395,共3页
Shanxi Medical Journal
关键词
线粒体脑肌病
DNA
线粒体
分子诊断技术
Mitochondrial encephalomyopathies
DNA,mitochondrial
Molecular diagnotic techniques
