Mitokondriyal Sitopati Öntanılı Hastalarda MT-TL1 Gen Mutasyonlarının İncelenmesi
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Date
2018Author
BULDUK, Bengisu Kevser
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Point
mutations in mitochondrial tRNA and rRNA genes cause oxidative phosphorylation
defects by negatively affecting protein synthesis function. The majority of
“mitochondrial cytopathies” are caused by point mutations in the tRNALeu (UUR) (MTTL1) gene. m.3243A>G mutation in the MT-TL1 gene is described in 80% of MELAS
patients. However, other rare mutations such as m.3256 C>T, m.3260 A>G, m.3271
T>C, m. 3291 T>C and m.3303 C>T are also identified in the same gene and associated
with mitochondrial cytopathies. Within the scope of this study, a novel ARMS-PCR
assay has been standardized to screen for the six described point mutations in the MTTL1 gene. The descriptive sensitivity of this approach was aimed to be improved by
combining it with HRM technology. Detection limits and the precision of this novel
method is verified by using recombinant mtDNA standards carrying heteroplasmic
mutations at different rates which is created by PCR-based mutagenesis. Peripheral
blood or skeletal muscle tissue samples of 219 patients, prediagnosed as mitochondrial
cytopathies at the Departments of Neurology and Child Neurology Units of Hacettepe
University Hospitals were screened using the developed method. The results of the
screening procedure revealed two patients to have m.3243 A> G mutation, one patient
with m.3290 T> C variation, and one patient carrying m.3303 C>T mutation. These
mutations were further confirmed by DNA sequence analysis. This novel ARMS-PCR
assay study and its results provide a new and technological "proof of concept" for
future screening of other rarely observed mtDNA variations in patients.