Comprehensive Scanning of the Entire Mitochondrial Genome for Mutations
| العنوان: | Comprehensive Scanning of the Entire Mitochondrial Genome for Mutations |
|---|---|
| المؤلفون: | Haeyoung Kwon, Lee-Jun C. Wong, Ren-Kui Bai, Duan-Jun Tan, John Park, Min-Hui Liang |
| المصدر: | Clinical Chemistry. 48:1901-1912 |
| بيانات النشر: | Oxford University Press (OUP), 2002. |
| سنة النشر: | 2002 |
| مصطلحات موضوعية: | Genetics, Homoplasmy, Mitochondrial DNA, Mutation, Mitochondrial Diseases, Mitochondrial disease, Biochemistry (medical), Clinical Biochemistry, Biology, medicine.disease, medicine.disease_cause, DNA, Mitochondrial, Polymerase Chain Reaction, Genome, Heteroplasmy, Mitochondria, medicine, Humans, Electrophoresis, Polyacrylamide Gel, Female, Restriction fragment length polymorphism, Child, Gene, Polymorphism, Restriction Fragment Length |
| الوصف: | Background: Definitive molecular diagnosis of mitochondrial disorders has been greatly hindered by the tremendous clinical and genetic heterogeneity, the heteroplasmic condition of pathogenic mutations, and the presence of numerous homoplasmic mitochondrial DNA (mtDNA) variations with unknown significance. We used temporal temperature gradient gel electrophoresis (TTGE) to detect heteroplasmic mutations from homoplasmic variations in the whole mitochondrial genome. Methods: We screened 179 unrelated patients by TTGE with use of 32 overlapping primer pairs. Mutations were identified by direct sequencing of the PCR products and confirmed by PCR with allele-specific oligonucleotide or restriction fragment length polymorphism analysis. Results: We detected 71 heteroplasmic and 647 homoplasmic banding patterns. Sequencing of the heteroplasmic fragments identified 68 distinct novel mutations and 132 reported sequence variations and mutations; most of them occurred only once. The deleterious nature of some of the novel mutations was established by analyzing the asymptomatic family members and the biochemical and molecular characteristics of the mutation. When the number of mutations was normalized to the size of the region, the occurrence of mutations was 2.4 times more frequent in the tRNA genes than in the mRNA (protein coding) regions. Conclusions: Screening by TTGE detects low proportions of mutant mtDNA and distinguishes heteroplasmic from homoplasmic variations. Results from comprehensive molecular analysis should be followed up with clinical correlation to establish a guideline for complete mutational analysis of the entire mitochondrial genome and to facilitate the diagnosis of mitochondrial disorders. |
| تدمد: | 1530-8561 0009-9147 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::40cace39dec6d4a02421696d018081ac https://doi.org/10.1093/clinchem/48.11.1901 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....40cace39dec6d4a02421696d018081ac |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15308561 00099147 |
|---|