【摘要】 目的:建立嗜肺军团菌mip基因荧光定量PCR检测方法。方法:利用Primer Express软件设计特异性引物和探针,对质粒标准品、嗜肺军团菌、非嗜肺军团菌及其他菌株进行检测,验证方法的特异性、敏感性和重复性,最后采集环境样本进行检测。结果:该方法特异性好,嗜肺军团菌呈现阳性结果,而非嗜肺军团菌及其他菌株均为阴性结果。检测灵敏度达293copies/μl;重复性较好,Ct值变异系数较小;检测的12份环境样本中5份阳性。结论:该方法快速且具有较好的特异性、敏感性、重复性,适于外环境嗜肺军团菌污染调查及应急事件的快速检测。更多还原

【Abstract】 Objective:To establish a fluorescence quantitative PCR assay to detection Legionella pneumophila.Methods:Primers and probe were designed by Primer software,and plasmid standard,legionella pneumophila,non-legionella pneumophila and other strains were used to evaluate the specificity,sensitivity and repeatability of the assay.Environmental samples were collected and detected by this assay.Results:This assay had high specificity for detecting Legionella pneumophila but not to non-legionella pneumophila and other strains.The sensitivity of this assay was 293 copies/μl.The repeatability of this assay was high,the coesfficient of variation of Ct values was low.Five of twelves environmental samples were positive.Conclusion:The fluorescence quantitative PCR provides a specific,rapid and sensitive method for quantitative detection of Legionella pneumophila.It is helpful for the rapid detection of environment source of Legionella pollution and emergency.更多还原