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1995 Fiscal Year Final Research Report Summary

Identification of periodontal pathogen by DNA probe-biolobiclal and chemical luminescence system

Research Project

Project/Area Number 05557086
Research Category

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

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

Principal Investigator

ABIKO Yoshimitsu  NIHON UNIVERSITY,SCHOOL OF DENTISTRY AT MATSUDO,PROFESSOR, 松戸歯学部, 教授 (70050086)

Co-Investigator(Kenkyū-buntansha) OSHIHARA Wataru  TORAY INDUSTRIES,INC.BASIC RESERACH LABORATORIES,, 研究員
KIYAMA Michiko  NIHON UNIVERSITY,SCHOOL OF DENTISTRY AT MATSUDO,, 松戸歯学部, 副手 (50256905)
HAYAKAWA Mitsuo  NIHON UNIVERSITY,SCHOOL OF DENTISTRY AT MATSUDO,, 松戸歯学部, 講師 (10112955)
Project Period (FY) 1993 – 1995
Keywordsperiodontal disease / bacterial examination / P.gingivalis / A.actinomycetemcomitans / A.viscosus. / DNA probe / riboprobe / genetic diagnosis
Research Abstract

One approach to the development of a rapid, sensitive and specific assay for the detection of the pathogens is molecular hybridization by using DNA or RNA probe. We focused to detection and identification of periodontal pathogens, P.gingivalis and A.actinomycetemcomitans as disease-active site and A.viscosus as inactive site during the progression of periodontal disease. We cloned genes ; outer membrane protein and glycylprolyl amonopeptidase from P.gingivalis, leukotoxin from A.actinomycetemcomitans and sialidase from A.viscosus. Then non-radiolabeled riboprobes were developed using in vitro transcription RNA synthesizing system. All probes were highly specific for identification of each pathogen.
We also examined the usefullness of alkaline phosphatase/lumigen PPD detection system using anti-DNA-RNA complex antibody. Result showed a good corelation between optical density and number of molecules until 10^4 molecules but poor corelation was found over 10^5 molecules. These unfortunate results may occur by contamination of RNA or DNA-RNA hybrid in the sample. Then we amplified the specific region from chromosomal DNA and measured by the same procedure, this experiment produced a good corelation for detection of pathogens.

  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] Yoshimitsu ABIKO,Kunihiro UEDA,Hiroshi NOJIMA,Youji MURAYAMA,and Sumiharu NOJI: "Passibilities of Genetic Diagnosis for Periodontal Disease" Dental Outlook. Vol.82. 41-54 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto TSUSHIMA,Mitsuo HAYAKAWA and Hisashi TAKIGUCHI: "Gene Cloning of the Specific Region of Porphyromonas gingivalis 40-kDa Outer Membrane Protein Gene and Development of Non-radioactive DNA Probe" Nihon University Journal of Oral Science. Vol.20. 29-39 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hideki TAMAYA,Koichi HIRATSUKA and Hisashi TAKIGUCHI: "Development of the Synthetic Porphyromonas gingivalis Outer Membrane Protein Gene Oligonucleotide DNA Probe" Nihon University Journal of Oral Science. Vol.20. 406-413 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sumiharu NOJI,Hidemi KURIHARA,Yoshimitsu ABIKO,and Youji MURAYAMA: "Prospect for Genetic (DNA) Diagnosis for dental Field" The Journal of the Japan Dental Association. Vol.47. 17-27 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshimitsu ABIKO: "Progress of Genetic Diagnosis in Future ; Utilization of DNA Diagnosis for Bacterial Examination in Periodontal Disease" Dental Outlook. Vol.84. 840-851 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto KOITABASHI,Mitsuo HAYAKAWA and Hisashi TAKIGUCHI: "Development of Porphyromonas gingivalis Glycylprolyl Aminopeptidase Riboprobe" Nihon University Journal of Oral Science. Vol.20. 499-507 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masataka ARAYA,Michiko KIYAMA and Hisashi TAKIGUCHI: "Development of Actinobacillus actinomycetemcomitans Leukotoxin Gene Riboprobe" Nihon University Journal of Oral Science. Vol.22. 27-35 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Katsukata SHIMIZU,Mitsuo HAYAKAWA and Hisashi TAKIGUCHI: "Construction of the Gene Clone Highly Expressing Porphyromonas gingivalis Glycylprolyl Diaminopeptidase" Nihon University Journal of Oral Science. Vol.22. 36-44 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Koichi HIRATSUKA,Mitsuo HAYAKAWA,Hisashi TAKIGUCHI and Yoshimitsu ABIKO: "Characterization of recombinant 150-kDa S-layr protein from Campyrobacter rectus" Nihon University Journal of Oral Science. Vol.22 (in press). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Noriaki KANASUGI,Koichi HIRATSUKA,Mitsuo HAYAKAWA,Takuji IKEMI,Yuji NAMIKI,Hisashi TAKIGUCHI and Yoshimitsu ABIKO: "Detection of Porphyromonas gingivalis by DNA-RNA hybridization with non-radioactivity labeled riboprobe" Nihon University Journal of Oral Science. Vol.22 (in press). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshimitsu ABIKO,Mitsuo HAYAKAWA,Koichi HIRATSUKA,Yasuko SHIBATA,Michiko KIYAMA,Shigeno SAITO and Hisashi TAKIGUCHI: "Identification of periodontal diseases associated microorganisms using nonradioactive riboprobe" Dental Outlook. (in press). (1996)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1997-03-04  

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