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DNAチップを用いた口腔微生物叢の網羅的定量的プロファイリング

Research Project

Project/Area Number 15659500
Research Category

Grant-in-Aid for Exploratory Research

Allocation TypeSingle-year Grants
Research Field Social dentistry
Research InstitutionTohoku University

Principal Investigator

高橋 信博  東北大学, 大学院・歯学研究科, 教授 (60183852)

Co-Investigator(Kenkyū-buntansha) 小関 健由  東北大学, 大学院・歯学研究科, 教授 (80291128)
竹原 直道  九州歯科大学, 歯学部, 教授 (00038879)
佐藤 拓一  東北大学, 大学院・歯学研究科, 講師 (10303132)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2003: ¥2,700,000 (Direct Cost: ¥2,700,000)
KeywordsDNAチップ / 16S ribosomal RNA / DNAプローブ / DNAプライマー / 口腔細菌 / DNAプライヤー / 培養法 / PCR-RFLP法 / 菌種特異的PCR法
Research Abstract

本年度は、1.前年度設計した口腔細菌特異的プローブを作成し、DNAチップへの搭載の可否(実用性)を検討し、2.DNAチップへの搭載が可能と考えられた24種のプローブを用いて口腔微生物叢解析用DNAチップを試作し、口腔微生物種特異性、使用法などを検討した。
1.DNAチップ搭載用口腔細菌特異的プローブの作成:
プローブの特異性についてはGeneBankなどの既存のDNAデータベースから、プローブのDNAチップへの搭載の可否についてはDNAチップ作成手順などの実用性から検討し、その結果、24種の口腔微生物に対する特異的プローブが完成した。さらに、この1年間で新たに入手した口腔微生物16S ribosomal RNA遺伝子配列データに基づき、新たなプローブを設計・作成中である。
2.DNAチップ搭載用の24種の口腔微生物特異的プローブを用いた口腔微生物叢解析用DNAチップの試作:
DNAチップ作成共同研究企業にて、24種の口腔微生物特異的プローブを搭載したDNAチップを試作した。各プローブに該当する標準菌株由来DNAを用いて検討したところ、試作したDNAチップは24種の標準菌体由来DNAを特異的に検出することができることがわかった。さらに、DNAチップ上のスポットは肉眼で明瞭に判別でき、市販の可視光用透過型スキャナーで計測・記録することが十分可能であることが分かった。
現在、実際の臨床サンプルを用いて検討中であり、臨床サンプルに含まれる24種の口腔微生物の従来法による検出(通常のnested PCR法での検出)を終え、DNAチップによる検出との照合を進めつつある。

Report

(2 results)
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (35 results)

All 2005 2004 Other

All Journal Article (6 results) Publications (29 results)

  • [Journal Article] Expression of type IV collagen and laminin at the interface between epithelial cells and fibroblasts from human periodontal ligament.2005

    • Author(s)
      Shimonishi M
    • Journal Title

      Eur J Oral Sci 113(1)

      Pages: 34-40

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Detection frequency of periodontitis-associated bacteria by polymerase chain reaction in subgingival and supragingival plaque of periodontitis and healthy subjects.2004

    • Author(s)
      Mayanagi G
    • Journal Title

      Oral Microbiol Immunol 19(6)

      Pages: 379-385

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A method for mapping the distribution pattern of cariogenic streptococci within dental plaque in vivo.2004

    • Author(s)
      Kato K
    • Journal Title

      Caries Res 38(5)

      Pages: 448-453

    • Related Report
      2004 Annual Research Report
  • [Journal Article] PTC gene mutation and expression of SHH, PTC, SMO, and GLI-1 in odontogenic keratocysts.2004

    • Author(s)
      Ohki K
    • Journal Title

      Int J Oral Maxillofac Surg 33(6)

      Pages: 584-592

    • Related Report
      2004 Annual Research Report
  • [Journal Article] p53 gene status and expression of p53, MDM2, and p14ARF proteins in ameloblastomas.2004

    • Author(s)
      Kumamoto H
    • Journal Title

      J Oral Pathol Med 33(5)

      Pages: 292-299

    • Related Report
      2004 Annual Research Report
  • [Journal Article] K-Ras gene status and expression of Ras/mitogen-activated protein kinase (MAPK) signaling molecules in ameloblastomas.2004

    • Author(s)
      Kumamoto H
    • Journal Title

      J Oral Pathol Med 33(6)

      Pages: 360-367

    • Related Report
      2004 Annual Research Report
  • [Publications] Nobuhiro Takahashi: "Acid-neutralizing activity during amino acid fermentation by Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum."Oral Microbiology and Immunology. 18(2). 109-113 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Takuichi Sato: "Nested PCR for detection of mutans streptococci in dental plaque."Letters in Applied Microbiology. 37(1). 66-69 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Takuichi Sato: "Identification of mutans streptococci by restriction fragment length polymorphism analysis of polymerase chain reaction-amplified 16S ribosomal RNA genes."Oral Microbiology and Immunology. 18(5). 323-326 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Shoko Takahashi-Abbe: "Biochemical and functional properties of pyruvate formate-lyase (PFL)-activating system in Streptococcus mutans."Oral Microbiology and Immunology. 18(5). 293-297 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Harumi Miyasawa: "Xylitol inhibition on anaerobic acid production by Streptococcus mutans at various pH levels."Oral Microbiology and Immunology. 18(4). 215-219 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Junko Matsuyama: "Fermentation of five sucrose isomers by human dental plaque bacteria."Caries Research. 37(6). 410-415 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hatsue Kakuta: "Xylitol inhibition on acid-production and growth of mutans streptococci in the presence_of_various_dietary_sugars under strictly anaerobic conditions."Caries Research. 37(6). 404-409 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kazuo Kato: "A method for mapping the distribution pattern of cariogenic streptococci within dental plaque in vivo."Caries Research. 38(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kousuke Ohki: "PTC gene mutation and expression of SHH, PTC, SMO, and GLI-1 in odontogenic keratocysts."International Journal of Oral and Maxillofacial Surgery. 33(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hiroyuki Tada: "Proteolysis of ICAM-1 on human oral epithelial cells by cysteine proteinases (Gingipains) from Porphyromonas gingivalis leading to down-regulation of neutrophil adhesion."Journal of Dental Research. 82. B-41 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Takuichi Sato: "Combination of genus-specific PCR and RFLP analysis for identification of oral streptococci."Journal of Dental Research. 82. B-80 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Gen Mayanagi: "Detection of 25 periodontopathic bacteria from periodontal sites by nested PCR."Journal of Dental Research. 82. B-153 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Mitsuru Shimonishi: "Expression of type IV collagen and laminin in the interface between epithelial cells and fibroblasts from human periodontal ligament."Journal of Dental Research. 82. B-287 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nobuhiro Takahashi: "Metabolic pathway for palatinose of oral Actinomyces species."Journal of Dental Research. 82. B-360 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kazuo Kato: "Distribution profiles of cariogenic streptococci within dental plaque treated with stannous fluoride gel."Caries Research. 37(4). 267-268 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Jumpei Washio: "Hydrogen sulfide-producing bacreria in tongue coating and oral malodor."Journal of Dental Research. 83(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Mitsuru Shimonishi: "Expression of amelogenin in cultired epithelial rests of Malassez."Journal of Dental Research. 83(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Juta Sato: "Expression of non-collagenous bone proteins at epithelial cell-fibroblast interface."Journal of Dental Research. 83(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] 熊谷 崇: "歯垢中のmutans streptococciのPCR法による検出について"東北大学歯学雑誌. 22(1). 54 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 多田浩之: "Porphyromonas gingivalisジンジパインはヒト菌肉線維芽細胞CD14を分解してLPS不応答性を誘導する"日本細菌学雑誌. 58(1). 158 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 佐藤廉也: "矯正用アンカープレート周囲溝内の細菌叢のプロファイリング"歯科基礎医学会雑誌. 45(5). 271 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 佐藤拓一: "菌属特異的なPCRとRFLP解析による口腔レンサ球菌種の同定"歯科基礎医学会雑誌. 45(5). 271 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 大島勇人: "抗菌性薬剤に対するラット臼歯感染歯髄の反応"歯科基礎医学会雑誌. 45(5). 287 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 阿部昌子: "ミュータンスレンサ球菌乳酸脱水素酵素の活性調節に対する環境pHの影響"歯科基礎医学会雑誌. 45(5). 325 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 角田初恵: "キシリトールによるStreptococcus mutans増殖抑制効果"歯科基礎医学会雑誌. 45(5). 326 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 堀はるみ: "Streptococcus mutans酸産生能のキシリトール感受性の多様性とその生化学的機構"歯科基礎医学会雑誌. 45(5). 326 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 加藤一夫: "real-time定量PCRを利用した歯垢内細菌の層別密度分布の分析"口腔衛生学会雑誌. 53(4). 340 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 鷲尾純平: "口臭と舌苔中の総菌数および硫化水素産生菌数との関係"口腔衛生学会雑誌. 53(4). 344 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 五十嵐公英: "小児から高齢者に至る広範囲年齢層プラーク酸産生能のチェアーサイドでの評価"口腔衛生学会雑誌. 53(4). 381 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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