复合探针荧光定量PCR检测土拉弗朗西斯菌

Detection of Francisella tularensis using real-time fluorescent PCR with complex probe

目的:建立检测土拉弗朗西斯菌复合探针荧光定量PCR方法。方法:对土拉弗朗西斯菌的FopA基因设计引物和探针,用荧光定量PCR检测,并对特异性、灵敏度和重复性进行评价。结果:本方法对所试验的土拉弗朗西斯菌纯培养物的最低检测浓度为100 CFU/ml,定量的检测变异系数小于5%。在模拟的污染血液样品中的检出限为1×103 CFU/ml。结论:本文建立的复合探针荧光定量PCR方法能高效、敏感、特异地检测土拉弗朗西斯菌。

Objective:To develop an assay using real-time fluorescent PCR with complex probe for the detection of Francisella tularensis.Methods: The real-time fluorescent quantitative PCR assay for Francisella tularensis was developed,with the primers designed based on FopA gene,and the specificity and sensitivity were also analyzed.Results: The detection limit of the method was 100 CFU/ml and that of artificially contaminated blood sample was 1×103 CFU/ml.The coefficients of variations were less than 5%.Conclusion: The established complex probe real-time fluorescent quantitative PCR assay for Francisella tularensis is an effective method with good specificity and sensitivity.