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The establishment of clinical examination system to detect periodontal microorganism using specific DNA probes

Research Project

Project/Area Number 02454439
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Conservative dentistry
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

HIROFUJI Takao  Kyushu Univ., Fac. Dent., Assistant Professor, 歯学部, 講師 (10189897)

Co-Investigator(Kenkyū-buntansha) KABASHIMA Hiroaki  Kyushu Univ., Fac. Dent., Assistant, 歯学部, 助手 (20214504)
HAMACHI Takafumi  Kyushu Univ., Fac. Dent., Assistant, 歯学部, 助手 (80198811)
MAEDA Katsumasa  Kyushu Univ., Fac. Dent., Professor, 歯学部, 教授 (00117243)
岩本 恭行  九州大学, 歯学部, 助手 (20203416)
青野 正男  九州大学, 歯学部, 教授 (70037498)
Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1992: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1990: ¥4,600,000 (Direct Cost: ¥4,600,000)
KeywordsPeriodontitis / Porphyromonas gingivalis / Cysteine proteinase / DNA cloning / Actinobacillus actinomycetemcomitans / Synthetic oligonucleotide / PCR method / Porphyromonas gingivalis / 多形核白血球機能抑制因子 / DNA遺伝子クロ-ニング / PCR / <Bacteroides>___ー <gingivalis>___ー / 歯周炎関連細菌由来特異的DNA遺伝子
Research Abstract

Prophyromonas gingivalis (P.g.) has been implicated as a key organism in 2dult periodontitis. A cysteine proteinase was purified from the culture supernatant of P.g. to a homogeneity by several continuous chromatographic methods. The molecular size of this enzyme was estimated to be 50KDa and the pl of this factor was between pH 4.5 to 5.1. This proteinase was found to induce the dysfunction of polymorphonuclear leukocytes (PMNs), as judged from the suppression of their chemiluminescence response. These results indicate that P.g. release the unique cysteine proteinase having a damaging effect on the host defense system. We are now considering the possibility that this factor has been developed to aid in the diagnosis and treatment of human periodontal disease using DNA technology. Actinobacillus actinomycetemcomitans (A.a.) is generally recognized as the main infectious organism in localized juvenile periodontitis and also found in adult periodontitis patients. We prepared for several synthetic olygonucleotides derived from IktA gene in A.a., then, used as primers of polymerase chain reaction method (PCR method). One of these primers reacted specifically all genomes in A.a. we tested by PCR method and amplified only one specific band on DNA gel electrophoresis. When this primer was reacted with samples from subgingival plaques of twenty one patients with periodontal periodontitis, samples from six-teen patients with periodontitis were positive by PCR method and these reactions were specific.
These results indicate that this PCR method using specific synthetic oligonucleotide as a primer is very useful to detect A.a. in subgingival site in patient with periodontitis.

Report

(4 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (1 results)

All Other

All Publications (1 results)

  • [Publications] M.Yoneda: "Suppression of bactericidal activity of human polymorphonuclear leukocytes by <Bacteroides>___ー <gingivalis>___ー" Infection and Immunity. 58. 406-411 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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