2001 Fiscal Year Final Research Report Summary
DNA DIAGNOSIS OF RARE GENETIC DISEASES USING JAPANESE MICROARRAY
| Project/Area Number |
12557224
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Human genetics
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
MATSUBARA Yoichi TOHOKU UNIV, MEDICAL GENETICS, PROFESSOR, 大学院・医学系研究科, 教授 (00209602)
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| Co-Investigator(Kenkyū-buntansha) |
KURE Shigeo TOHOKU UNIV, MEDICAL GENETICS, ASSOCIATE PROFESSOR, 大学院・医学系研究科, 助教授 (10205221)
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| Project Period (FY) |
2000 – 2001
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| Keywords | DNADIAGNOSIS / RARE GENETIC DISEASES / SINGLE-GENE DISORDER / PHENYLKETONURIA / GLYCOGEN STORAGE DISEASE |
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| Research Abstract |
The rapidly growing mutation database raises the notion of a DNA-based diagnosis of various genetic diseases by screening known mutations. A mutation prevalent in a defined population is a good candidate for this type of approach. Within this context, it is important to develop a robust DNA diagnostic method that is suitable for clinical application. In the current study, we have explored the possibility of applying a DNA microarray method for the simultaneous detection of multiple disease-causing mutations. The method we tested was mini-sequencing employing fluorescence-labeled dNTP. DNA fragments containing mutation-sites were amplified by multiplex PCR, denatured, and hybridized with oligonucleotides immobilized on a glass plate. Primer extension reaction was performed in the presence of Cy5-labeled dNTP. After washing, the array was scanned with laser-beam to detect incorporated fluorescent signals. The poor signal-to-noise ratio in this method, however, hampered reproducible and reliable genotyping. We therefore applied a new genotyping method "DNA stick" for the detection of various mutations. Successful genotyping indicated that this method may be readily applied to a microarray format with further modification. In addition, we were able to identify prevalent genetic mutations in glycogen storage disease type la and type lb, hereditary deafness due to connexin26 mutations, and holocarboxylase synthetase deficiency to aid developing a mutation-panel useful in Japanese population.
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