CAJ | 학술논문

目的：建立针对嗜肺军团菌Mip基因的实时荧光定量TaqMan PCR检测方法，并进行自来水和空调冷却水模拟标本的检测评价。方法：根据嗜肺军团菌Mip基因的特异性序列设计引物和TaqMan探针，建立嗜肺军团菌的实时荧光定量TaqMan PCR快速检测方法，对方法进行灵敏度及特异性评价，并对自来水和空调冷却水模拟标本中的嗜肺军团菌进行检测。结果：建立的方法对嗜肺军团菌的检测具有高度特异性，与3种非嗜肺军团菌和6种其他呼吸道病原均没有交叉反应；基因组DNA的检测灵敏度为1.6 pg/μL，模拟自来水和空调冷却水标本的检测灵敏度为10 CFU/mL。结论：建立的TaqMan荧光定量PCR方法特异、灵敏、快速，适于嗜肺军团菌的日常监测和暴发疫情的应急诊断。
목적：건립침대기폐군단균Mip기인적실시형광정량TaqMan PCR검측방법，병진행자래수화공조냉각수모의표본적검측평개。방법：근거기폐군단균Mip기인적특이성서렬설계인물화TaqMan탐침，건립기폐군단균적실시형광정량TaqMan PCR쾌속검측방법，대방법진행령민도급특이성평개，병대자래수화공조냉각수모의표본중적기폐군단균진행검측。결과：건립적방법대기폐군단균적검측구유고도특이성，여3충비기폐군단균화6충기타호흡도병원균몰유교차반응；기인조DNA적검측령민도위1.6 pg/μL，모의자래수화공조냉각수표본적검측령민도위10 CFU/mL。결론：건립적TaqMan형광정량PCR방법특이、령민、쾌속，괄우기폐군단균적일상감측화폭발역정적응급진단。
Objective: To develop a TaqMan-based real-time PCR method for rapid detection of Legionella pneu-mophila. Methods: The sequence of macrophage infectivity potentiator(Mip) gene was downloaded from GenBank and the specific primers and TaqMan probe were designed in the conserved region of the Mip gene for L.pneu-mophila. Then, the real-time PCR array for rapid detection of L.pneumophila was developed and its spceificity and sensitivity were evaluated. Simulated environment water samples were used to assess the assay. Results: Only L. pneumophila strains generated fluorescent signals, and no cross-reaction was observed for the differential control strains including three non-pneumophila strains and six other respiratory pathogens. The detection limits were 1.6 pg/μL with genomic DNA of L.pneumophila, and 10 CFU/mL with simulated water samples. Conclusion: The Taq-Man real-time PCR assay described here is specific, sensitive and rapid for detection of L.pneumophila, and this assay could be used for laboratory-based monitoring and emergency detection of L.pneumophila.