Precise and simultaneous quantification of mitochondrial DNA heteroplasmy and copy number by digital PCR.

ABSTRACT: Mitochondrial DNA (mtDNA) is present in multiple copies and phenotypic consequences of mtDNA mutations depend on the mutant load surpassing a specific threshold. Additionally, changes in mtDNA copy number can impact mitochondrial ATP production, resulting in disease. Therefore, the precise determination of mtDNA heteroplasmy and copy number is crucial to the study of mitochondrial diseases. However, current methods can be imprecise, and quantifying small changes in either heteroplasmy or copy number is challenging. We developed a new approach to measure mtDNA heteroplasmy using a single digital PCR (dPCR) probe. This method is based on the observation that fluorescent-labeled probes in dPCR exhibit different intensities depending on the presence of a single nucleotide change in the sequenc

SUBMITTER: Shoop WK 

PROVIDER: S-EPMC9650046 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature