| Project/Area Number |
24659285
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | Kanazawa Medical University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
BABA Hisashi 金沢医科大学, 医学部, 准教授 (60359750)
SUZUKI Masahiro 愛知県衛生研究所, 生物学部, 研究員 (70446649)
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2012: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | 緑膿菌 / 遺伝子タイピング / PCR-based ORF typing法 / MLST / POT法 |
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| Research Abstract |
We developed a novel digital genotyping method of Pseudomonas aeruginosa (P. aeruginosa PCR-based ORF typing method; POT typing method). According to the distribution of the ORF (Open reading frame) in the clinical isolates, 10 ORF originated from genomic islet and 5 ORF originated from genomic island to increase the discriminatory power were selected. Metallo-beta-lactamase (MBL) (IMP or VIM group) gene and P. aeruginosa marker gene were added for the typing, finally the primers were adapted for the 10 and 9-plex PCR. The PCR results were scored with a 1 or 0 according to the presence or absence of the amplicon of the selected genes in order of primer number, and the resulting binary code was transformed into decimal numbers, separating into two parts with 10 (genomic islet, POT1) and 9 (genomic island and MBL, POT2) figures. POT1 was shown to be related with Multilocus Sequence Typing (MLST) number, and the discriminatory power was equivalent to PFGE.
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