發(fā)布時(shí)間：2018-09-04 07:54

【摘要】：建立了毛細(xì)管電泳快速檢測牙齦卟啉單胞菌(P.g)、齒垢密螺旋體(T.d)、福賽斯坦納菌(T.f)3種牙周病病原菌種的方法。紫外分光光度法表明,采用無菌吸潮紙尖及PBS緩沖液可以實(shí)現(xiàn)牙周病原菌的快速有效提取。對(duì)提取的牙周病病原菌種做多聚酶鏈?zhǔn)椒磻?yīng)(PCR)與多重PCR反應(yīng),最后以20 cm長,75μm內(nèi)徑的石英毛細(xì)管作為分離通道,分離電壓4000 V,1.2%羥乙基纖維素(HEC,250 K)為篩分介質(zhì),牙周病病原菌菌種P.g,T.d,T.f PCR及多重PCR產(chǎn)物12 min內(nèi)得到較好分離。結(jié)果表明,毛細(xì)管電泳與PCR技術(shù)相結(jié)合,可以實(shí)現(xiàn)牙周病原菌種快速鑒定,且檢測限低至4.80×10-11ng/μL。方法已用于牙周病原菌菌種的快速檢測。
[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.
【作者單位】： 上海理工大學(xué)光電信息與計(jì)算機(jī)工程學(xué)院;大阪大學(xué)應(yīng)用物理系;
【基金】：國家自然科學(xué)基金(21205078) 高等學(xué)校博士學(xué)科點(diǎn)專項(xiàng)科研基金博導(dǎo)類資助課題(20123120110002) 上海市教育發(fā)展基金會(huì)曙光計(jì)劃項(xiàng)目(11SG44) 上海高校青年教師培養(yǎng)資助計(jì)劃(51-13-302-102)資助
【分類號(hào)】：R781.4;O658.9

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