PCR-SSCP在检测Leber遗传性视神经病变线粒体DNA突变的应用

Application of PCR-SSCP in detecting mtDNA Mutation for the patients with Leber hereditary optic neuropathy

目的探讨PCR-SSCP技术检测Leber遗传性视神经病变(LHON)线粒体DNA(mtDNA)3个原发突变的最佳分析条件。方法应用PCR-SSCP技术检测LHONmtDNA,对主要影响SSCP分辨率的聚丙酰胺凝胶的浓度和组成优化分析,并与突变特异性引物PCR(MSP)、限制性片段长度多态性(FRLP)及测序结果相印证。结果80g/L交联度为66:1的非变性聚丙酰胺凝胶能同时检出LHONmtDNA的三个原发致病突变,与MSP、FRLP及DNA测序的结果一致。结论该分析条件简便、快速,能有效地检测LHONmtDNA突变。

Objective: To explore the best analyzed condition of PCR-SSCP in detecting three mtDNA mutation for the patients with Leber hereditary optic neuropathy(LHON).Methods:PCR-SSCP was used to detect mtDNA untation and three groups of deferent components of polyacrylamide gel were used to select the best one for further analysis.The results were compared with PCR(MSP),FRLP and DNA sequence methods.Results:Our results show that the best condition is 80g/L non-denaturing polyacrylamide gel(Acr:Bis,66∶1)that can produce clear bands and detect three primary mutation of LHON mtDNA,and is similar to the results of MSP,FRLP and DNA sequence methods.Conclusion:The analyzed condition in our study is simple,quick and can effect to detect LHON mtDNA mutation.