Development of PCR based open reading frame typing method for Clostridium difficile
| Project/Area Number |
15K08663
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Fujita Health University (2017)
Aichi Prefectural Institute of Public Health (2015-2016) |
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Principal Investigator |
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| Research Collaborator |
YAMADA Kazuhiro 愛知県衛生研究所, 生物学部, 主任 (90564857)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | Clostridioides difficile / 分子疫学 / POT法 / Clostridium difficile / 分子疫学解析 / PCR / PCR-ribotyping / MLST |
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| Outline of Final Research Achievements |
Novel molecular epidemiology method called PCR-based ORF typing (POT) was developed for Clostridium difficile isolates. The genotype of POT is determined by detecting ORFs with plus, minus manner. ORFs choosing in this study shows different distribution patterns depending on genetic background of isolates. The ORFs were selected by comparing whole genome data of C. difficile, and testing clinical isolates using PCR. Finally, 10 ORFs showing correlations to MLST, and 6 ORFs which can classify isolates with same ST were selected. POT was constructed from 11-plex PCR detecting the 10 ORF and C. difficile marker, and 10-plex PCR detecting the 6 ORFs, toxins (toxin A, B, and binary toxin), and C. difficile marker. The discrimination power demonstrated by Simpson’s index was 0.976. POT can perform within 4 hours using conventional PCR apparatus and get easy understandable genotypes. POT will make molecular epidemiological analysis of C. difficile easy and contributes to infection control.
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Report
(4 results)
Research Products
(12 results)
