全唾液中变异链球菌受体的检测

Detection of receptors of Streptococcus mutants in human whole salivary components

目的通过变异链球菌黏结素蛋白和唾液蛋白的相互作用,筛选全唾液中变异链球菌黏结素蛋白的受体成分。方法变异链球菌国际标准菌株经活化、液体培养,采用冷冻乙醇法提取在液体培养时细菌分泌至培养液中的黏结素,并对黏结素的成分进行电泳分析、含量测定和抗原性检测。选取15名高龋者和15名无龋者,分别于进食后2 h采集其无刺激性全唾液,经冷冻干燥后制备电泳样品,采用SDS聚丙烯酰胺凝胶电泳(SDS-PAGE)将其分开,用变异链球菌黏结素蛋白和变异链球菌的抗体成分进行Western-blotting免疫印迹检测。结果在细菌培养液中提取到了变异链球菌表面分泌的黏结素成分。免疫印迹检测结果显示,唾液中变异链球菌的受体成分至少有6种,不只局限于目前认识的富脯蛋白等3种。结论变异链球菌表面黏结素可与全唾液中的多种蛋白结合,这有可能是变链菌黏附于牙齿表面的物质基础。

Objective To select the acceptor ingredients of Streptococcus mutans protein in the whole saliva through the interaction between adhesion and saliva protein of Streptococcus mutans.Methods After activation and liquid culture of Streptococcus mutans strains of international standards,we took Su-bonding from liquid culture of bacteria secretion in the process of liquid culture by using chilled ethanol extraction.Then su-bonded components underwent electrophoresis analysis,content determination and antigen detection.We collected 15 with high carries and 15 with no carries.Two hours after eating,the whole non-irritating saliva was collected,electrophoresis samples were prepared after freeze-drying,and then stains were separated with SDSPAGE.Western blot detection was used for adhesive protein and Streptococcus mutans＇ antibody composition.Results The ingredients of Su-bonding secreted on the surface of Streptococcus mutans were obtained in the cultured saliva.There were at least six kinds of components in the extraction of Western blot detection,not limited to the three known kinds of proline-rich protein.Conclusion The adhesion of Streptococcus mutants can be bound with the proteins of saliva.It may be the material basis in Streptococcus mutants adhered to tooth surface.