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Mitochondrial and nuclear disease panel (Mito-aND-Panel): Combined sequencing of mitochondrial and nuclear DNA by a cost-effective and sensitive NGS-based method.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Abicht, Angela 
Scharf, Florentine 
Kleinle, Stephanie 
Schön, Ulrike 
Holinski-Feder, Elke 

Abstract

BACKGROUND: 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. METHOD: 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 (WES). RESULTS: The Mito-aND-Panel permits accurate detection of low-level mtDNA heteroplasmy due to a very high sequencing depth compared to standard diagnostic procedures using Sanger sequencing/SNaPshot and WES which is crucial to identify maternally inherited mitochondrial disorders. CONCLUSION: We established a NGS-based method with combined sequencing of the complete mtDNA and nuclear genes which enables a more sensitive heteroplasmy detection of mtDNA mutations compared to traditional methods. 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.

Description

Keywords

NUMTs, heteroplasmy detection, high-throughput sequencing, mitochondrial and nuclear disease panel, mitochondrial disorders, mtDNA-server, Costs and Cost Analysis, DNA, Mitochondrial, Genetic Testing, High-Throughput Nucleotide Sequencing, Humans, Mitochondrial Diseases, Sensitivity and Specificity, Sequence Analysis, DNA

Journal Title

Mol Genet Genomic Med

Conference Name

Journal ISSN

2324-9269
2324-9269

Volume Title

6

Publisher

Wiley
Sponsorship
Medical Research Council (MR/N025431/2)
Wellcome Trust (109915_A_15_Z)