Studies on etiology of periodontal disease and its diagnosis and therapy

从8例牙周炎患者的13个牙周袋中分离出188株牙龈卟啉单胞菌，以FunA基因为基础进行限制性片段长度多态性(RFLP)分析。共有5种RFLP基因分型，其中一半以上的菌株属于I型。4例患者仅携带1种RFLP基因，1例患者在不同部位仅携带2种RFLP基因。另一方面，在另外3例患者中，在单个牙周袋中发现多种RFLP基因。牙周病患者牙周菌斑中分离到90株临床分离的口腔密螺旋体，是否多种基因同时定植于一个牙周袋，还是发生了突变，有待进一步研究。用生物化学方法、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和DNA斑点杂交法进行了细胞酶、蛋白质分析。在基本无血清的10(M10)培养基上分离到60株细菌。其余的在含血清的培养液中分离。根据菌株的生化特性，将其分为6个组。6个组中有3个组为无糖组，其中2个组为齿密螺旋体和“温州密螺旋体”。其余3组均为糖解组，分为9个亚组。通过与苏氏密螺旋体DNA探针的细胞酶、细胞蛋白和斑点杂交分析表明，所有的糖解基团均为苏氏密螺旋体或其亲缘关系较近的种。这项研究表明，从人牙周袋中提取的M10可以鉴定出新发现的糖解型口腔密螺旋体。

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.