Chip-based mtDNA mutation screening enables fast and reliable genetic diagnosis of OXPHOS patients.
OBJECTIVE
Oxidative phosphorylation is under dual genetic control of the nuclear and the mitochondrial DNA (mtDNA). Oxidative phosphorylation disorders are clinically and genetically heterogeneous, which makes it difficult to determine the genetic defect, and symptom-based protocols which link clinical symptoms directly to a specific gene or mtDNA mutation are falling short. Moreover, approximately 25% of the pediatric patients with oxidative phosphorylation disorders is estimated to have mutations in the mtDNA and a standard screening approach for common mutations and deletions will only explain part of these cases. Therefore, we tested a new CHIP-based screening method for the mtDNA.


METHODS
MitoChip (Affymetrix) resequencing was performed on three test samples and on 28 patient samples.


RESULTS
Call rates were 94% on average and heteroplasmy detection levels varied from 5-50%. A genetic diagnosis can be made in almost one-quarter of the patients at a potential output of 8 complete mtDNA sequences every 4 days. Moreover, a number of potentially pathogenic unclassified variants (UV) were detected.


CONCLUSIONS
The availability of long-range PCR protocols and the predominance of single nucleotide substitutions in the mtDNA make the resequencing CHIP a very fast and reliable method to screen the complete mtDNA for mutations.
van Eijsden
Rudy G E
RG
Department of Clinical Genetics, Maastricht University, Maastricht, The Netherlands.
Gerards
Mike
M
Eijssen
Lars M T
LM
Hendrickx
Alexandra T M
AT
Jongbloed
Roselie J E
RJ
Wokke
John H J
JH
Hintzen
Rogier Q
RQ
Rubio-Gozalbo
Maria E
ME
De Coo
Irenaeus F M
IF
Briem
Egill
E
Tiranti
Valeria
V
Smeets
Hubert J M
HJ
D002648:Child; D004252:DNA Mutational Analysis / Q000379:methods*; D004272:DNA, Mitochondrial / Q000032:analysis* / Q000235:genetics; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D028361:Mitochondrial Diseases / Q000175:diagnosis*; D009154:Mutation; D009690:Nucleic Acid Conformation; D020411:Oligonucleotide Array Sequence Analysis; D010085:Oxidative Phosphorylation; D016133:Polymerase Chain Reaction; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics
D004272:DNA, Mitochondrial; D012343:RNA, Transfer