毛细管电泳技术快速检测牙周病原菌

发布时间：2018-09-04 07:54

【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。
[Abstract]:A capillary electrophoresis (CE) method for the rapid detection of three kinds of periodontal pathogenic bacteria, P.gingivalis (P.G), T. denticulata (T.d) and T.Forsestana (T.F), was established. Ultraviolet spectrophotometry showed that the rapid and effective extraction of periodontal pathogenic bacteria could be achieved by using aseptic hygroscopic paper tip and PBS buffer. Polymerase chain reaction (PCR) with multiple PCR was performed on the isolated periodontal pathogenic bacteria. Finally, quartz capillary with a diameter of 75 渭 m long for 20 cm was used as the separation channel, and the separation voltage was 4000 V / L 1.2% hydroxyethyl cellulose (HEC,250 K) as the screening medium. The pathogen of periodontal disease was isolated from T. ddtf PCR and multiple PCR products in 12 min. The results showed that the rapid identification of periodontal pathogenic bacteria could be achieved by combining capillary electrophoresis with PCR technique, and the detection limit was as low as 4.80 脳 10-11ng/ 渭 L. The method has been used for rapid detection of periodontal pathogenic bacteria.
【作者单位】：上海理工大学光电信息与计算机工程学院;大阪大学应用物理系;
【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
【分类号】：R781.4;O658.9

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