NCM 2021: Mitochondrial DNA analysis by long-read NGS in patients affected by mitochondrial disease

Human mitochondria produce ATP and metabolites to support development and maintain cellular homeostasis. These organelles possess their own genome (mtDNA) and defects in its replication can cause mtDNA structural alterations, leading to early death, neuromuscular and neurodegenerative diseases, or cancer. Short-read NGS is the standard approach for the analysis of mtDNA; however, intrinsic limitations hamper the detection of mtDNA structural alterations. In this study, we tested and optimized nanopore long-read NGS for the sequencing of mtDNA from muscle biopsy of patients affected by mitochondrial diseases, in order to overcome short-read limitations and increase the accuracy for the identification of mtDNA large deletions and complex re-arrangements.