Gharbia S E, Haapasalo M, Shah H N, Kotiranta A, Lounatmaa K, Pearce M A, Devine D A
Faculty of Dentistry, Dalhousie University, Halifax, Nova Scotia.
J Periodontol. 1994 Jan;65(1):56-61. doi: 10.1902/jop.1994.65.1.56.
The occurrence and surface properties of prevotella intermedia and P. nigrescens in healthy sites and in periodontic and endodontic infections were studied among 73 strains, tentatively identified as P. intermedia. Fifteen strains were from necrotic root canal infections, 41 were from periodontal samples, and 17 isolates were obtained from healthy gingival sites. Identification of isolates as either P. intermedia or P. nigrescens was based on differences in malate and glutamate dehydrogenase electrophoretic mobilities which allowed unambiguous separation of P. intermedia and P. nigrescens. The majority of strains from periodontal samples were P. intermedia (29 of 41 strains). In endodontic samples only 4 out of 15 isolates were P. intermedia, while all except 1 of 17 strains from healthy gingival sites were identified as P. nigrescens. SDS-PAGE of whole cell proteins revealed 31 and 38 kDa proteins in P. nigrescens which were not detected in P. intermedia. Surface biotinylation of cells, followed by Western blotting and detection by alkaline phosphatase conjugated extravidin, showed strong staining of the 31 kDa protein in P. nigrescens indicating that this protein is located on the surface of the cell. Corresponding staining was not seen in P. intermedia. Fimbria-like projections were observed using electron microscopy of negatively-stained cells of P. nigrescens. The results show that P. intermedia and P. nigrescens may have different site specificities and surface properties and thus emphasize the need for accurate identification of these two species for the evaluation of their role in the pathogenesis of oral infections.
在73株初步鉴定为中间普氏菌的菌株中,研究了中间普氏菌和变黑普氏菌在健康部位、牙周感染和牙髓感染中的发生情况及表面特性。15株来自坏死根管感染,41株来自牙周样本,17株分离自健康牙龈部位。根据苹果酸和谷氨酸脱氢酶电泳迁移率的差异将分离株鉴定为中间普氏菌或变黑普氏菌,这使得中间普氏菌和变黑普氏菌能够明确区分。牙周样本中的大多数菌株为中间普氏菌(41株中的29株)。在牙髓样本中,15株分离株中只有4株是中间普氏菌,而来自健康牙龈部位的17株菌株中,除1株外均被鉴定为变黑普氏菌。全细胞蛋白的SDS-PAGE显示,变黑普氏菌中有31 kDa和38 kDa的蛋白,而在中间普氏菌中未检测到。细胞表面生物素化,然后进行蛋白质印迹并用碱性磷酸酶偶联抗生物素蛋白检测,结果显示变黑普氏菌中31 kDa蛋白有强烈染色,表明该蛋白位于细胞表面。在中间普氏菌中未观察到相应的染色。使用变黑普氏菌负染细胞的电子显微镜观察到菌毛样突起。结果表明,中间普氏菌和变黑普氏菌可能具有不同的位点特异性和表面特性,因此强调了准确鉴定这两个菌种对于评估它们在口腔感染发病机制中作用的必要性。