Minisequencing mitochondrial DNA pathogenic mutations

BMC Med Genet. 2008 Apr 10:9:26. doi: 10.1186/1471-2350-9-26.


Background: There are a number of well-known mutations responsible of common mitochondrial DNA (mtDNA) diseases. In order to overcome technical problems related to the analysis of complete mtDNA genomes, a variety of different techniques have been proposed that allow the screening of coding region pathogenic mutations.

Methods: We here propose a minisequencing assay for the analysis of mtDNA mutations. In a single reaction, we interrogate a total of 25 pathogenic mutations distributed all around the whole mtDNA genome in a sample of patients suspected for mtDNA disease.

Results: We have detected 11 causal homoplasmic mutations in patients suspected for Leber disease, which were further confirmed by standard automatic sequencing. Mutations m.11778G>A and m.14484T>C occur at higher frequency than expected by change in the Galician (northwest Spain) patients carrying haplogroup J lineages (Fisher's Exact test, P-value < 0.01). The assay performs well in mixture experiments of wild:mutant DNAs that emulate heteroplasmic conditions in mtDNA diseases.

Conclusion: We here developed a minisequencing genotyping method for the screening of the most common pathogenic mtDNA mutations which is simple, fast, and low-cost. The technique is robust and reproducible and can easily be implemented in standard clinical laboratories.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Mutational Analysis / methods*
  • DNA Primers
  • DNA, Mitochondrial / genetics*
  • Humans
  • Leigh Disease / genetics
  • MELAS Syndrome / genetics
  • Optic Atrophy, Hereditary, Leber / genetics*
  • Point Mutation*
  • Polymerase Chain Reaction
  • Polymorphism, Single Nucleotide
  • Spain


  • DNA Primers
  • DNA, Mitochondrial