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Studies on etiology of periodontal disease and its diagnosis and therapy

牙周病的病因及诊治研究

基本信息

批准号：
05454515
负责人：
ISHIKAWA Isao
金额：
$ 4.35万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

A total of 188 porphyromonas gingivalis strains isolated from 13 periodontal pockets of 8 periodontitis patients were investigated by means of restriction fragment length polymorphism (RFLP) analysis using the funA gene as a pobe. A total of 5 RFLP genotypes were identified, and over half of the isolates belonged to type I.Four of the 8 patients harbored only 1 RFLP genotype of P.gingivalis, and 1 patient harbored only 2 RFLP genotypes in different sites. On the other hand, in 3 other patients, multiple RFLP genotypes were found in single periodontal pockets. Further studies will be required to clarify whether multiple genotypes of P.gingivalis colonized a single periodontal pocket simultaneously or whether a mutation occurred in the fimbrilin gene locus of P.gingivalis.A total of 90 clinical strains of oral treponemes was isolated from subgingivalplaque in patients with periodontal disease. They were characterized by biochemical means as well as cell enzyme, protein analisys using sodium dodecyl sulfatepolyacrylamide gel electrophoresis and DNA dot blot hybridization. Sixty strains were isolated on Medium 10 (M10), which was fundamentally serum-free. The remainder were isolated on serum-containing media. Isolates were divided into 6 groups according to their biochemical characteristics. Three of the 6 groups were asaccharolytic, and 2 of these 3 groups were Treponema denticola and "Treponema vincentii". The other 3 groups were saccharolytic and further divided into 9 subgroups. The analyzes of cell enzyme, cell protein and dot blot hybridization with the DNA probe of Treponema socranskii indicated that all the saccharolytic groups were T.socranskii or closely related species. This study indicated that the newly characterized saccharolytic oral treponemes could be identified using M10 from the human periodontal pocket.

以funA基因为探针，通过限制性片段长度多态性（RFLP）分析，对从8名牙周炎患者的13个牙周袋中分离出的总共188株牙龈卟啉单胞菌进行了研究。共鉴定出 5 种 RFLP 基因型，其中超过一半属于 I 型。8 名患者中，4 名仅携带 1 种牙龈卟啉单胞菌 RFLP 基因型，1 名患者在不同部位仅携带 2 种 RFLP 基因型。另一方面，在另外 3 名患者的单个牙周袋中发现了多种 RFLP 基因型。是否多个基因型的牙龈卟啉单胞菌同时定殖在单个牙周袋中，或者是否在牙龈卟啉单胞菌的菌毛蛋白基因位点发生突变，还需要进一步的研究。从牙周病患者的龈下斑块中分离出总共90株口腔密螺旋体临床菌株。通过生化方法以及细胞酶、十二烷基硫酸钠聚丙烯酰胺凝胶电泳和DNA点印迹杂交的蛋白质分析对它们进行表征。在基本无血清的培养基 10 (M10) 上分离出 60 株菌株。剩余物在含血清培养基上分离。根据其生化特征，分离株分为6组。 6组中的3组是解糖菌，这3组中的2组是齿垢密螺旋体和“文森密螺旋体”。其余3组为糖分解组，进一步分为9个亚组。细胞酶、细胞蛋白分析及与索克兰斯基螺旋体DNA探针的斑点杂交分析表明，所有解糖类群均为索克兰斯基螺旋体或近缘种。这项研究表明，可以使用来自人类牙周袋的 M10 来鉴定新鉴定的解糖口腔密螺旋体。