Rapid identification of mitochondrial DNA (mtDNA) mutations in neuromuscular disorders by using surveyor strategy

Department of Medical Genetics, Archet 2 Hospital, CHU Nice, France.
Mitochondrion (Impact Factor: 3.25). 04/2008; 8(2):136-45. DOI: 10.1016/j.mito.2007.10.008
Source: PubMed


Mutations of mitochondrial genome are responsible for respiratory chain defects in numerous patients. We have used a strategy, based on the use of a mismatch-specific DNA endonuclease named " Surveyor Nuclease", for screening the entire mtDNA in a group of 50 patients with neuromuscular features, suggesting a respiratory chain dysfunction. We identified mtDNA mutations in 20% of patients (10/50). Among the identified mutations, four are not found in any mitochondrial database and have not been reported previously. We also confirm that mtDNA polymorphisms are frequently found in a heteroplasmic state (15 different polymorphisms were identified among which five were novel).

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    • "Janne et al. has applied this platform for high-sensitivity mutation screening of the epidermal growth factor receptor (EGRF) gene in human cancer specimens [10], but it has not been applied for the detection of mtDNA mutation. Although Bannwarth et al. have developed 17 primer pairs for screening mutations of entire human mtDNA by using SN analysis, they detected cleaved DNA fragments by using agarose gel electrophoresis [11], [12]. Because SN/WAVE-HS analysis does not require the testing on optimal temperatures and can be used to analyze amplicons much longer than 500 bp, it is very likely to be a more superior technique than DHPLC for the efficient screening of unknown heteroplasmic variants or mutations in entire mtDNA. "
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