| Co-Investigator(Kenkyū-buntansha) |
ITO Katsutoshi Showa university, Pharmaceutical sciences, Lecture, 薬学部, 講師 (20223141)
MAEDA Masako Showa university, Pharmaceutical sciences, Professor, 薬学部, 教授 (00053869)
|
|---|
| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥2,500,000 (Direct Cost: ¥2,500,000)
|
|---|
| Research Abstract |
Gene diagnosis is utilized as diagnoses such as hereditary disease and infectious disease, and as diagnoses method such as the prediction of drug metabolic ability. Generally the molecular biological technique utilizing in the laboratory is used as DNA analysis of gene diagnosis. However, the rapid and high sensitive DNA analysis has been required in infectious disease and tailor made medicine. Recently, as the method, DNA chip method is developed, and the use in the clinical field is being tried. In this study, microchip electrophoresis was developed as high sensitive and rapid DNA analytical method, and the establishment of gene diagnosis which analyze DNA polymorphism with the one point mutation, O157 ( intestinal hemorrhagic Escherichia coli ) and mutans streptococcus was carried out. Construction of micro chip and selection of the polymer which is used as the separation medium were carried out. As the result, it was possible to analyze the PCR product of the O157 verotoxin gene (
… More
VT1 and VT2 ) using the channel of 30 μm depth and 100 μm width at separation length 1mm in slight10 seconds. And, in order to detect the one point mutation in the ras gene which is one of the oncogenes, the PCR was done mutation site, and next, SSCP (single-strand conformation polymorphism) analysis was carried out. In this study, separation media such as polymer and gel were mainly examined. As the result, the analysis of single nucleotide polymorphism (SNP) was difficult, though it was able to separate in the difference of large loop structure. Next, DNA diagnosis method of mutans streptococcus which existed in dental plaque, was developed as a diagnosis method of the dental treatment and care. Duplex allele specific PCR and common PCR for S. mutans and S. sobrinus were developed. And the obtained PCR products were analyzed by the imcrochip electrophoresis. By this method, these genes could be analyzed in 3 minutes. And, the detection limit was high sensitive of 20 fg. By the research of the above, the microchip electrophoresis seems to be the usefulness as a DNA detection method of the infectious disease which requires speedy DNA analysis. Less
|
