Mitochondrial and Nuclear Disease (Mito-aND-Panel): Combined Sequencing of Mitochondrial and Nuclear DNA by a Cost Effective and Sensitive NGS-based Method

The diagnosis of mitochondrial disorders is challenging because of the clinical variability and genetic heterogeneity of these conditions. „Next Generation Sequencing“ (NGS) technology offers a robust high-throughput platform for nuclear and mitochondrial DNA (mtDNA) analyses. We developed a custom Agilent SureSelect Mitochondrial and Nuclear Disease-Panel (Mito-aND-Panel) capture kit that allows parallel enrichment for subsequent NGS-based sequence analysis of nuclear mitochondrial disease-related genes and the complete mtDNA genome. Sequencing of enriched mtDNA simultaneously with nuclear genes was compared with the separated sequencing of the mitochondrial genome and whole exome sequencing. The „Mito-aND-Panel“ permits accurate detection of low-level mtDNA heteroplasmy due to a very high sequencing depth compared to traditional methods which is crucial to identify maternally inherited mitochondrial disorders. Because the method promotes the analysis of mtDNA variants in large cohorts, it is cost-effective and simple to setup, we anticipate this is a highly relevant method for sequence-based genetic diagnosis in clinical diagnostic applications.