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建立了毛细管电泳快速检测牙龈卟啉单胞菌(P.g)、齿垢密螺旋体(T.d)、福赛斯坦纳菌(T.f)3种牙周病病原菌种的方法。紫外分光光度法表明,采用无菌吸潮纸尖及PBS缓冲液可以实现牙周病原菌的快速有效提取。对提取的牙周病病原菌种做多聚酶链式反应(PCR)与多重PCR反应,最后以20 cm长,75μm内径的石英毛细管作为分离通道,分离电压4000 V,1.2%羟乙基纤维素(HEC,250 K)为筛分介质,牙周病病原菌菌种P.g,T.d,T.f PCR及多重PCR产物12 min内得到较好分离。结果表明,毛细管电泳与PCR技术相结合,可以实现牙周病原菌种快速鉴定,且检测限低至4.80×10-11ng/μL。方法已用于牙周病原菌菌种的快速检测。
A rapid capillary electrophoresis method was developed for the detection of three periodontal pathogens, P.g, T.d, and T.f. UV spectrophotometry shows that the use of sterile absorbency paper tip and PBS buffer solution can be realized quickly and effectively periodontal pathogens extraction. Polymerase chain reaction (PCR) and multiplex PCR were carried out on the extracted pathogenic bacteria of periodontal disease. Finally, a 20 cm long and 75 μm ID quartz capillary tube was used as the separation channel. The separation voltage was 4000 V and 1.2% hydroxyethylcellulose (HEC , 250 K) as screening medium, Pg, Td, Tf PCR and multiple PCR products of periodontal disease pathogen were separated within 12 min. The results showed that the combination of capillary electrophoresis and PCR could achieve rapid identification of periodontal pathogens and the detection limit was as low as 4.80 × 10-11 ng / μL. Method has been used for rapid detection of periodontal pathogens.