Triplex real-time PCR--an improved method to detect a wide spectrum of mitochondrial DNA deletions in single cells.

ABSTRACT: Mitochondrial DNA (mtDNA) mutations are commonly found in the skeletal muscle of patients with mitochondrial disease, inflammatory myopathies and sarcopenia. The majority of these mutations are mtDNA deletions, which accumulate to high levels in individual muscle fibres causing a respiratory defect. Most mtDNA deletions are major arc deletions with breakpoints located between the origin of light strand (OL) and heavy strand (OH) replication within the major arc. However, under certain disease conditions, rarer, minor arc deletions are detected. Currently, there are few techniques which would allow the detection and quantification of both types of mtDNA deletions in single muscle fibres. We have designed a novel triplex real-time PCR assay which simultaneously amplifies the MT-ND4 gene in t

SUBMITTER: Rygiel KA 

PROVIDER: S-EPMC4437295 | biostudies-literature | 2015 May

REPOSITORIES: biostudies-literature