Minisequencing mitochondrial DNA pathogenic mutations

作者：Alvarez Iglesias Vanesa; Barros Francisco; Carracedo Angel; Salas Antonio^*

来源：BMC Medical Genetics, 2008, 9(1): 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.

• 出版日期2008-4-10
• 单位河北医科大学