| Project/Area Number |
12557238
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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 |
Laboratory medicine
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| Research Institution | Gifu University |
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Principal Investigator |
EZAKI Takayuki Gifu Univ., Microbiology, Professor, 医学部, 教授 (90151977)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAMURA Yoshiaki Gifu Univ., Microbiology, Assist. Professor, 医学部, 助教授 (80262757)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥11,200,000 (Direct Cost: ¥11,200,000)
Fiscal Year 2001: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2000: ¥9,300,000 (Direct Cost: ¥9,300,000)
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| Keywords | Lebel 2 / Lebel 3 / detection / DNA chip / Identification / Microarray / Multiplex PR / Diagnosis / 感染症診断 / 病原細菌 / 感染症 / 診断 |
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| Research Abstract |
Human pathogenic bacteria are classified as level 2 ( 350 species ) and level 3 ( 33 species ). We prepared ribosomal RNA microarray for most of these level 2 and 3 pathogens to establish comprehensive identification and detection system of pathogenic bacteria. PCR products of ribosomal DNA of these were quantitatively Spotted on glass slide. Cye3 or Cye5 labeled PCR products from unidentified bacterial colonies were hybridized On DNAs of slides and hybridized DNAs were quantitatively measured by a laser scanner. Ribosomal RNA array could not differentiate some potent pathogens which shared similar ribosomal RNA sequences. Therefore, we prepared pathogenic factprs DNA microarray for them.
Immobilizing these two different sources of PCR products, we could identify and detect most of human pathogenic level 2 and level 3 pathogens. Universal primers were not applicable to amplify contaminated sample such as stool, sputum and urinary smears. So we prepared each specific primer and mixed in a single tube to simplify amplification procedure. This method successfully worked. We mixed more than 50-100 different specific primers but could successfully amplify DNA of pathogens.
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