Molecular Epidemiological Study of Periodontopathic Bacteria in Periodontal Diseases

牙周病牙周病菌的分子流行病学研究

• 批准号: 11672082
• 负责人: KOKEGUCHI Susumu
• 金额: $ 2.3万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11672082/
• 关键词: Periodontal disease; Porphyromanas gingivalis; Actinobacillub actinomycetemcomitans; Infection; Transmission; Porphyromanas gingivalis

Previous epidemiological studies of Porphyromonas gingivalis(Pg)and Actinobacillus actinomycetemcomitans(Aa)in periodontal diseases have performed to analyze their clinical isolates from diseased periodontal regions by biotyping, serotyping, ribotyping and genotyping. These typing methods following by their bacterial culture and isolation are time-consuming and complicated. It is difficult to fully elucidate the microflora of individulas with periodontals diseases by analysing the limited number of clinical isolates. The aim of this study was to examine the direct identification of Pg and Aa in subgingival palque samples and further their genotyping based on the specific antigen genes amplified by polymerase chain reaction(PCR). In this study, the outer membrane protein(OMP)gene and fimbrial protein(FMP)gene were selected as the specific antigen gene for Pg and Aa, respectively. Our PCR method for specific antigen genes could allow the direct and specific detection of Pg and Aa in plaque samples. Pg were classified into two OMP genotypes(pga53 and pga67)by PCR and Aa were differenciated into four FMP genotypes by PCR-restriction fragment-length polymorphism(RFLP). We examined the distribution and prevalence of Pg and Aa genotypes in 59 patients(145 sites)with periodontal diseases. Almost of the patients were infected with only single genotype of Pg and/or Aa. In the case of two families(mother-son-daughter), the identical genotypes of Pg and Aa were distributed among family members. These results indicated that periodontal diseases might be initiated and progressed by clonal genotype of Pg and/or Aa, which might be possibly transmitted within family. The PCR-genotyping method based on specific antigen genes of Pg and Aa would be useful tools for their molecular epidemiological study in periodontal diseases and provide new insights into their transmission and pathogenesis.

以往对牙周病中的牙龈卟啉单胞菌（Pg）和放线菌（Aa）进行了流行病学研究，通过生物分型、血清分型、核糖分型和基因分型分析了它们在牙周病变区域的临床分离株。这些分型方法，然后是细菌培养和分离，耗时且复杂。通过分析有限数量的临床分离株，很难完全阐明牙周病患者的微生物群。本研究的目的是检验龈下牙菌斑样品中Pg和Aa的直接鉴定，并基于聚合酶链反应（PCR）扩增的特异性抗原基因进一步进行基因分型。本研究选择外膜蛋白（OMP）基因和毛膜蛋白（FMP）基因分别作为Pg和Aa的特异性抗原基因。我们的特异性抗原基因PCR方法可以直接特异检测斑块样品中的Pg和Aa。通过PCR将Pg分为2种OMP基因型（pga53和pga67），通过PCR-限制性片段长度多态性（RFLP）将Aa分为4种FMP基因型。我们检测了59例牙周病患者（145个部位）Pg和Aa基因型的分布和流行情况。几乎所有患者仅感染单一基因型的Pg和/或Aa。在两个家庭（母子女）中，Pg和Aa基因型在家庭成员中分布相同。这些结果表明牙周病可能是由Pg和/或Aa克隆基因型引发和发展的，并可能在家族内传播。基于Pg和Aa特异性抗原基因的pcr基因分型方法，将为其在牙周病中的分子流行病学研究提供有用的工具，并为其传播和发病机制提供新的认识。

项目成果

Sawada S., Kokeguchi S., et al.: "Phylogenetic characterization of Centipeda periodontii, Selenomonas sputigena and Selenomonas species by 16S rRNA gene sequence analysis"Microbios. 133. 133-140 (1999)

Sawada S.、Kokeguchi S. 等人：“通过 16S rRNA 基因序列分析对 Centipeda periodontii、Selenomonas sputigena 和 Selenomonas 物种进行系统发育表征”Microbios。

Sawada S., Kokeguchi S., et al.: "Development of 16S rDNA-based PCR assay for detecting Centipeda periodontii and Selenomonas sputigena"Letters in Applied Microbiology. 30. 423-426 (2000)

Sawada S.、Kokeguchi S. 等人：“开发基于 16S rDNA 的 PCR 检测方法，用于检测牙周蜈蚣和痰硒单胞菌”应用微生物学快报。

Sawada S.,Kokeguchi S.,Takashiba S., et al.: "Development of 16S rDNA-based PCR assay for detecting Centipeda periodontii and Selenomonas sputigena"Letters in Applied Microbiology. 30(6). 423-426 (2000)

Sawada S.、Kokeguchi S.、Takashiba S. 等人：“开发基于 16S rDNA 的 PCR 检测方法，用于检测牙周蜈蚣和痰硒单胞菌”应用微生物学快报。

Sawada S., Kokeguchi S., et al.: "Phylogenetic characterization of Centipedapriodontii, Selenomonas sputigena and Selenomonas spp by 16Sr RNA gene sequence analysis"Microbios. 98. 133-140 (1999)

Sawada S.、Kokeguchi S. 等人：“通过 16Sr RNA 基因序列分析对 Centipedapriodontii、Selenomonas sputigena 和 Selenomonas spp 进行系统发育表征”Microbios。

本行博,苔口進ら: "2つの特異的外膜蛋白遺伝子を用いたParphyromonas gingivalisの型別とそれを基にした歯周ポケットにおける感染分布の研究"日本歯科保存学会雑誌. 42. 58 (1999)

Yukihiro Moto、Susumu Kokeguchi 等人：“使用两种特定外膜蛋白基因对牙龈卟啉单胞菌进行类型分类，并据此研究牙周袋中的感染分布”日本保守牙科学会杂志 42. 58 (1999)。