title: [Detection of mtDNA 11778 (G-->A) point mutation in a family with Leber's hereditary optic neuropathy by site-specific polymerase chain reaction].
abstract: OBJECTIVE
  To find a simple, fast, accurate, and quantitative PCR-based method for mutation detection, so as to identify mitochondrial DNA 11778 G-->A point mutation in patients with Leber's hereditary optic neuropathy (LHON).
  
  
  METHODS
  On the basis of sequencing of mtDNA from LHON proband, M primer for mutation and N primer for normal were designed to be coupled with reverse primer respectively. Specific PCRs were done on an amplifying condition with high stringency such as a well controlled annealing temperature, low Mg2+ concentration and less thermal cycles. The objective pedigree includes 10 individuals, were against 40 normal control persons.
  
  
  RESULTS
  Different ratios of indicative mtDNA 11778A-->G mutation were checked out from the proband, affected maternal members and a 10 year-old boy (up to now no appearance yet), whereas not appeared on normal spouses, paternal offsprings in the family, neither did on 40 controls.
  
  
  CONCLUSIONS
  This site-specific PCR method is a kind of general mutation analysis way, without the restriction of existence of endonuclease site. It can be applied for the gene diagnosis of known-mutation hereditary diseases such as LHON.
authors.affiliation: Central Laboratory, First Hospital of Peking University, Beijing 100034, China.
authors.forename: Shu-lan
authors.initials: SL
authors.lastname: Niu
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yu-feng
authors.initials: YF
authors.lastname: Xu
authors.forename: Ding-fang
authors.initials: DF
authors.lastname: Bu
authors.forename: Ze-qin
authors.initials: ZQ
authors.lastname: Ren
authors.forename: Shao-yu
authors.initials: SY
authors.lastname: Wang
authors.forename: Guo-hua
authors.initials: GH
authors.lastname: Liu
authors.forename: Yu
authors.initials: Y
authors.lastname: Qi
mesh_terms: D000328:Adult; D004272:DNA, Mitochondrial / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D029242:Optic Atrophy, Hereditary, Leber / Q000235:genetics*; D010375:Pedigree; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D004272:DNA, Mitochondrial