2型糖尿病患者线粒体常见基因突变PCR检测的局限性
Limitation on detecting common gene mutations of mitochondrial DNA in type 2 diabetes with PCR
-
摘要: 目的: 研究北京地区汉族人群中线粒体DNA(m itochondrial DNA,m tDNA)多位点突变频率与2型糖尿病之间的相关性。方法: 采用直接PCR、PCR-限制性片段长度多态性(restriction fragm ent length polymorph ism,RFLP)、PCR-时相温度梯度凝胶电泳(temporal temperature grad ient gel electrophoresis,TTGE)法筛查m tDNA 4977片段缺失、线粒体tRNALeu(UUR)A3243G突变和m tDNA T14577C突变。结果: 在250例2型糖尿病患者及142例健康对照外周血基因组总DNA中,未发现m tDNA 4977缺失、线粒体tRNALeu(UUR)A3243G突变及m tDNA T14577C突变。结论: mtDNA 4977缺失、线粒体tRNALeu(UUR)A3243G突变及m tDNA T14577C突变在外周血中没有检出,采用常规PCR法检测2型糖尿病患者外周血线粒体DNA突变具有较大的局限性。Abstract: Objective: To study the association of common gene mutations of mitochondrial DNA（mtDNA） in patients with type 2 diabetes mellitus（DM2）. Methods: Polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP） and Polymerase chain reaction-temporal temperature gradient gel electrophoresis（PCR-TTGE） were carried out to screen the fragment deletion of mtDNA 4977 and the mutations of tRNA Leu（UUR）A 3243G and mtDNA T14577C. Results: Fragment deletion of mtDNA 4977,mutations of tRNA Leu（UUR）A3243G and mtDNA T14577C were not found neither in DM2 nor healthy controls. Conclusion: There is a limitation on detecting common gene mutations of mitochondrial DNA in type 2 diabetes by PCR for the heterogenicity of mtDNA in the blood samples.
下载: