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

• DOI: 10.1016/j.jbc.2022.102574
• Publication Date: 2022-10-06T16:01:16Z
• Authors: Wendy K. Shoop; Cassandra L. Gorsuch; Sandra R. Bacman; Carlos T. Moraes
• Keywords: Heteroplasmy

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 copy number can impact mitochondrial ATP production, resulting disease. Therefore, precise determination heteroplasmy crucial to study diseases. However, current methods be imprecise, quantifying small either or challenging. We developed new approach measure using single digital PCR (dPCR) probe. This method based observation that fluorescent-labeled probes dPCR exhibit different intensities depending presence nucleotide change sequence bound by finding allowed us precisely simultaneously determine levels duplex dPCR. tested this two models (human mouse), which proved faster more internally controlled when compared other published routinely used genetics field. believe could broadly applicable detection quantification mixed genetic variations.