Studies on etiology of periodontal disease and its diagnosis and therapy

对8例牙周炎患者13个牙周袋中分离的188株牙龈卟啉单胞菌进行了限制性片段长度多态性（RFLP）分析，以funA基因为探针。共鉴定出5种RFLP基因型，半数以上的分离株属于i型。8例患者中4例仅携带1种RFLP基因型，1例患者在不同部位仅携带2种RFLP基因型。另一方面，3例患者在单个牙周袋中发现多个RFLP基因型。是否牙龈卟啉单胞菌的多个基因型同时定殖在单个牙周袋内，或者牙龈卟啉单胞菌的纤原蛋白基因位点是否发生了突变，还需要进一步的研究来阐明。从牙周病患者的龈下菌斑中分离出口腔密螺旋体临床菌株共90株。采用生化手段对其进行鉴定，并用十二烷基硫酸钠凝胶电泳和DNA斑点杂交对其进行酶学、蛋白分析。在基本无血清的Medium 10 （M10）培养基上分离到60株菌株。其余的在含血清培养基上分离。根据分离菌株的生化特性将其分为6组。6组中3组为无糖解菌，3组中2组为密螺旋体和文森密螺旋体。其余3组为糖溶性，又分为9个亚组。细胞酶学分析、细胞蛋白分析和DNA探针斑点杂交表明，所有溶糖类群均为索氏密螺旋体或近缘种。本研究表明，新鉴定的解糖性口腔密螺旋体可以用来自人牙周袋的M10进行鉴定。

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.