Detection of Leber hereditary optic neuropathy using single strand conformation polymorphism analysis

Objective To screen mtDNA mutations of Leber hereditary optic neuropathy,and to investigate the clinical characteristics and cell biological alternation of the disease.Methods Three pedigrees were inquired and 30 maternal members of them were tested.Following informed consent,periphery blood samples were obtained.Thirty-two healthy persons without optic disease were included in control group.Total DNA was isolated using routine way.Four mtDNA fragments were amplified using four pairs of primers,which covered eleven nucleotide position mutations reported abroad.Then the product was analysed by single strand conformation polymorphism analysis.Results There were 41 blood species members in the three pedigrees including 23 male and 19 female.Fifteen males and 5 females were affected.Compared with control group,single stranth conformation polymorphism analysis of polymerase chain reaction products amplified with primer pair 1 in the experiment group,showed bands shift in all lanes,only one lane showed band shift in products amplified with primer pair 2,and no band shift was found in products amplified with primer pair 3 and 4.Conclusion The age of onset was ahead of schedule with increasing passing generation within these pedigrees;Nucleotide position 11 778 mutation is overwhelming majority of Leber hereditary optic neuropathy patients in China;All materal members in these three pedigrees carry same mtDNA mutation.Only a part of them have been affected.Members with symptoms are Leber hereditary optic neuropathy patients and their asymptomatic maternal relatives are carriers.