嗜肺军团菌血清1型荧光PCR检测方法的研究  

Study on the Detection of Legionella pneumophila Serotype1 by Fluorescence PCR

作　　者：王仲敏 宋喆 王全新 王飞 王乃福 刘培 高旗利 WANG Zhong-Min;SONG Zhe;WANG Quan-Xin;WANG Fei;WANG Nai-Fu;LIU Pei;GAO Qi-Li(Animal,Plant and Foodstuff Inspection Center,Tianjin Customs,Tianjin 300041)

机构地区：[1]天津海关动植物与食品检测中心,天津300041

出　　处：《中国口岸科学技术》2023年第7期92-96,共5页China Port Science and Technology

基　　金：天津海关科研项目(2020THK007)。

摘　　要：本研究利用嗜肺军团菌O抗原基因簇wzt基因为靶基因,设计并筛选出一对实时荧光PCR引物和TaqMan探针,建立了嗜肺军团菌血清1型(Lp1)实时荧光PCR检测方法。特异性试验显示,3株Lp1出现了典型的扩增曲线,23株近源参考菌株没有扩增。灵敏度试验显示,水中Lp1的检测底线可达到1.1×10^(2) CFU/mL。重复性试验结果显示,高、中、低浓度扩增Ct值相对标准偏差分别为0.51%、0.42%、0.31%。结果表明,该方法特异性好、灵敏度高、重复性好、检测流程短,适于水环境污染状况调查及应急事件快速检测。According to the O-antigen gene cluster wzt gene of Legionella pneumophila,a pair of real-time PCR speciffc primers and TaqMan probe were designed and screened,and a real-time PCR method for the detection of Legionella pneumophila serotype1(Lp1)was established.The results of speciffcity test showed that 3 reference strains of Legionella pneumophila serotype1 had typical ampliffcation,and 23 strains of near source reference were not ampliffed.The sensitivity test showed that the detection limit of Legionella pneumophila serotype1 in water could reach 1.1×10^(2) CFU/mL.The repeatability test showed the RSD of ampliffcation Ct in different concentrations were 0.51%,0.42%,0.31%respectively.The method has good speciffcity and repeatability,high detection sensitivity,and short detection process,which is suitable for the investigation of water environment pollution and the rapid detection of emergency events.

关 键 词：嗜肺军团菌血清1型 wzt基因 实时荧光PCR 

分 类 号：O657.3[理学—分析化学] X832[理学—化学]