毛细管电泳技术快速检测牙周病原菌  

Rapid detection of periodontal pathogens by capillary electrophoresis

作　　者：李振庆[1] 孟凡明[1] 刘晨晨[1] 张大伟[1] 山口佳则[2] 

机构地区：[1]上海理工大学光电信息与计算机工程学院,上海200093 [2]大阪大学应用物理系

出　　处：《分析试验室》2014年第9期1009-1012,共4页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金(21205078);高等学校博士学科点专项科研基金博导类资助课题(20123120110002);上海市教育发展基金会曙光计划项目(11SG44);上海高校青年教师培养资助计划(51-13-302-102)资助

摘　　要：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。We developed a rapid way to detect periodontal pathogens （ e. g. , Porphyromonas gingivlis, （ P. g, ）, Treponema denticola, （T. d）, and Tannerela forsythia, （T. f） ） by capillary electrophoresis （CE） in this work. Ultraviolet-visible spectrophotometric method demonstrated that the periodontal pathogens could be effectively extracted by paper points and phosphate buffered saline （PBS） solution. Then we performed polymerase chain reaction （PCR） and muhi-PCR reaction of periodontal pathogens. Finally a coated silica capillary column （75 μm i.d × 20 cm） was employed as the separation channel, 4000 V as the separation voltage, 1.2% hydroxyethyl cellulose （250 K） as the separation polymer, the PCR and muhi-PCR products of P. g, T. d and T. f were effectively resolved within 12 min. Results show that CE conjunct with PCR can realize rapid detection of periodontal pathogens, and the limit of detection can reach as low as 4.80 × 10^ -11 ng/μL. Such a method is of great clinical significance for rapid detection of periodontal pathogens. Keywords： Capillary electrophoresis ; Periodontal pathogens ; Polymerase chain reaction ; DNA

关 键 词：毛细管电泳 牙周病原菌 聚合酶链式反应 脱氧核糖核酸 

分 类 号：O657.8[理学—分析化学]