Construction of the custom-made DNA chip for pathogenic diagnosis of bacteria related with periodontal disease
| Project/Area Number |
13671981
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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 |
病態科学系歯学(含放射線系歯学)
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| Research Institution | Nihon University |
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Principal Investigator |
HIRATSUKA Koichi Nihon University, School of Dentistry at Matsudo, Lecturer (Full-Time), 松戸歯学部, 講師 (80246917)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2003: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | periodontal disease / pathogen / microarray / transcriptome / examination / environment / Porphyromonas gingivalis / 口腔細菌 / ストレス / 診断 / P. gingivalis / 増殖 / 酸化ストレス / 遺伝子発現 / リアルタイムPCR / 蛍光標識 / Porphyromonas gingivalis / ヘミン / 解析 / チップ / クローニング |
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| Research Abstract |
It takes long time and much cost to analyze the expressions of all known-functional genes by the conventional method. Furthermore, we cannot expect to utilize any commercial-based microarrays for oral bacterial in the future. Therefore, we prepared a DNA custom-made array of pathogenicity-associated genes in Porphyromonas gingivalis, strongly associated with disease activity in adult periodontal disease, and established the microarray procedure such as hybridization, wash, and labeling condition, and how to analyze. Consequently, mRNA was good start material for catching low signal intensities of gene expressions, compares with Total RNA. Moreover, it became clear to match results between microarray and Real-time PCR when we used fixed quantity of the labeled external standards from human genes for chip-chip normalization rather than 16S rRNA (internal standard). Following various environmental change was stressed to P. gingivalis and the transcriptome profiling was analyzed (1. Gene expression profiling during growth; 2. Gene expression profiling under hemin limitation; 3. Gene expression profiling under oxidization stress). These results suggested that we can expect not only to detect gene expressions of various pathogen but also to demonstrate the relationship among the pathogenic genes in P. gingivalis. Moreover, this technology expected a possibility that it could monitor of the degree of activity of the bacteria in a periodontal pocket.
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Report
(4 results)
Research Products
(3 results)
