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A new method for analysis of mitochondrial DNA point mutations and assess levels of heteroplasmy

2006; Elsevier BV; Volume: 342; Issue: 2 Linguagem: Inglês

10.1016/j.bbrc.2006.01.152

1090-2104

Denise Cassandrini, Maria Grazia Calevo, Alessandra Tessa, Giovanni Manfredi, Fabiana Fattori, Maria Chiara Meschini, Rosalba Carrozzo, E. Tonoli, Marina Pedemonte, Carlo Minetti, Federico Zara, Filippo M. Santorelli, Claudio Bruno,

ATP Synthase and ATPases Research

Determination of mitochondrial DNA (mtDNA) heteroplasmy for the diagnosis of patients with mitochondrial disorders is a difficult task due to the coexistence of wild-type and mutant genomes. We have developed a new method for genotyping and quantification of heteroplasmic point mutations in mtDNA based on the SNaPshot technology. We compared the data of this method with the widely used "last hot-cycle" PCR-RFLP method by studying 15 patients carrying mtDNA mutations. We showed that SNaPshot is an accurate, reproducible, and sensitive technique for the determination of heteroplasmic mtDNA mutations in different tissues from patients, and it is a promising system to be used in prenatal and postnatal diagnosis of mtDNA-associated disorders.