Diagnosis of mitochondrial disorders applying massive pyrosequencing.
Mitochondrial disorders are a frequent cause of neurological disability affecting children and adults. Traditionally, molecular diagnosis of mitochondrial diseases was mostly accomplished by the use of Sanger sequencing and PCR-RFLP. However, there are particular drawbacks associated with the use of these methods. Recent multidisciplinary advances have led to new sequencing methods that may overcome these limitations. Our goal was to explore the use of a next generation sequencing platform in the molecular diagnosis of mitochondrial diseases reporting our findings in adult patients that present with a clinical-pathological diagnosis of a mitochondrial encephalomyopathy. Complete genomic sequences of mitochondrial DNA were obtained by 454 massive pyrosequencing from blood samples. The analysis of these sequences allowed us to identify two diagnostic pathogenic mutations and 74 homoplasmic polymorphisms, useful for obtaining high-resolution mitochondrial haplogroups. In summary, molecular diagnosis of mitochondrial disorders could be efficiently done from readily accessible samples, such as blood, with the use of a new sequencing platform.
Consultorio de Neurogenética, Centro Universitario de Neurología JM Ramos Mejía, Buenos Aires, Argentina. marcelokauffman@marcelokauffman.info
Marcelo Andrés
MA
Kauffman
Dolores
D
Gonzlez-Morón
Damián
D
Consalvo
Gastón
G
Westergaard
Martín
M
Vazquez
Estefanía
E
Mancini
Ana Lía
AL
Taratuto
Raúl
R
Rey
Silvia
S
Kochen
D000328:Adult; D004272:DNA, Mitochondrial / Q000235:genetics; D017809:Fatal Outcome; D005260:Female; D005819:Genetic Markers; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D028361:Mitochondrial Diseases / Q000175:diagnosis* / Q000235:genetics; D025202:Molecular Diagnostic Techniques; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D004272:DNA, Mitochondrial; D005819:Genetic Markers