Most pathogenic human mitochondrial DNA (mtDNA) mutations are Heteroplasmic and are difficult to detect when present at very low levels in a population of normal mtDNA molecules. This paper describes a simple methodology to detect low levels of the single base pair Heteroplasmic MELAS (A3243G) mutation. A series of peptide nucleic acids (PNAs) was designed to bind to the wild type mtDNA, thus reducing the PCR amplification of the wild type mtDNA to background levels while permitting the mutant DNA to become the dominant product and readily discernable. This methodology will permit easy detection of the MELAS A3243G mutation is asymptomatic or symptomatic carriers with low to undetectable blood levels of this mutation.
Citation: Clinical Chemistry
Pub Type: Journals
heteroplasmy, human, MELAS, mitochondrial DNA (mtDNA), peptide nucleic acid (PNA)