Use of single strand conformation polymorphism analysis to detect point mutations in human mitochondrial DNA

J Neurol Sci. 1992 Sep;111(2):222-6. doi: 10.1016/0022-510x(92)90074-u.

Abstract

Myoclonus epilepsy with ragged-red fibers (MERRF) has been shown to be associated with a specific point mutation at the nucleotide 8344 in the tRNA(Lys) gene of mitochondrial DNA (mtDNA). We screened 6 patients with clinically diagnosed MERRF and 1 patient with ocular myopathy for point mutations in the tRNA(Lys) gene, using single strand conformation polymorphism (SSCP) analysis, which can detect even a 1-basepair difference between 2 DNA sequences. Using SSCP and consequent DNA sequencing, we identified the known MERRF mutation in 4 out of 6 MERRF patients, as well as in 1 patient with a new clinical phenotype associated with this mutation: progressive external ophthalmoplegia, muscle weakness and a lipoma, but no myoclonus or epilepsy. Two of the patients with clinical MERRF had neither the MERRF-mutation nor any other mutations in the tRNA(Lys) gene. Using SSCP analysis, we also detected a new polymorphism in 1 patient. Thus, SSCP analysis can be applied to search effectively and rapidly for point mutations or polymorphisms in mitochondrial DNA.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • DNA Mutational Analysis*
  • DNA, Mitochondrial / genetics*
  • DNA, Single-Stranded / genetics
  • Female
  • Humans
  • MERRF Syndrome / genetics*
  • Male
  • Nucleic Acid Conformation
  • Nucleic Acid Renaturation
  • Polymerase Chain Reaction
  • RNA, Transfer, Lys / genetics*

Substances

  • DNA, Mitochondrial
  • DNA, Single-Stranded
  • RNA, Transfer, Lys