| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
Many human diseases are caused by small alterations in DNA sequence of the disease-causing genes. The alterations in DNA sequence include many types of mutation and polymorphism on disease-causing genes, such as one or several nucleotides replacement, deletion or insertion of some sequence, differences in a variable number of tandem repeat (VNTR) locus, and the instability of microsatellite repeat. DNA diagnosis for human diseases is, therefore, realized by the analysis of the alterations in the DNA sequence of disease-causing genes. In this study, we developed multicolor detectable laser-induced fluorescence (LIF) detection system for capillary electrophoresis and capillary array electrophoresis, and examined the efficiency and the performance of the systems coupled with polymerase chain reaction (PCR) technique for rapid genotyping of disease-causing genes and high-speed DNA diagnosis for human diseases. The system is successfully applied to rapid genotyping of apolipoprotein E gene (APOE) and p53 tumor suppressor gene through detection of one nucleotide replacement, apolipoprotein B gene (APOB) through typing of its VNTR locus, detection of an expansion for triplet repeat, and DNA mismatch repair gene through microsatellite instability analysis. Additionally, the systems are successfully applied to DNA diagnosis for cancer (p53 and DNA mismatch repair gene), Alzheimer's disease (APOE), coronary heart disease (APOB), and myotonic dystrophy (triplet repeat). In these instances, many data shown here demonstrate that capillary electrophoresis coupled with the PCR technique is a promising technology for DNA diagnosis of human diseases with high throughput, high accuracy, less labor-intensive, and cost effectiveness.
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