| Project/Area Number |
13670264
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | Kanazawa University |
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Principal Investigator |
FUJITA Shinichi Graduate School of Medical Science Kanazawa Uniersity Associate Professor, 大学院・医学系研究科, 助教授 (00115271)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Internal transcribed spacer / PCR / blood culture / rapid identification / microchip electrophoresis / Staphnlococcus aureus / Canodida / molecular technique / Jwternal Transcribed spacer / 同定 / 抗酸菌 / 真菌 / Clostridium difficile / ユニバーサルプライマー / マイクロチップゲル電気永動 / 泳動パターン / rRNA gene |
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| Research Abstract |
PCR amplification of the 16S-23S internal transcribed spacer (ITS) followed by microchip electerophoresis was evaluated for the usefulness in rapid identification of bacteria. Multiplex PCR amplification was used to test fungi, and Nocardia strains : the ITS 1 and 2 regions and the 5.8S rDNA region were amplified by using universal primers ITS1 and ITS4. The ITS2 region was simultaneously amplified by using universal primers ITS3 and ITS4.
ITS-PCR technique used in this study correctly identified clinically important bacterial strains, and Candida strains at species level within 2 h. Twenty-six different ITS-PCR patterns were demonstrated for 36 Clostridium difficile isolates on the basis of differences in the length of one or more amplicons. In addition, 202 Staphylococcal strains were divided into 40 ITS-PCR patterns. Therefore, the analysis of ITS-PCR patterns of C.difficile and staphylococci may be useful tool for epidemiological study of these strains.
The ITS-PCR method, prospectively applied to Gram staining-positive blood culture bottles, resulted in the correct detection and identification of bacterial strains and yeast strains. In conclusion, the ITS-PCR method followed by microchip electrophoresis seems to be a promising tool for the rapid identification of clinically important bacterial and yeast strains from culture colonies and from Gram staining-positive blood culture bottles.
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