实时荧光定量PCR法与常规PCR法及细菌培养法检测单增李斯特菌的比较

目的：比较实时荧光定量PCR法、常规PCR法及细菌培养法检测单核细胞增生性李斯特菌的灵敏度与特异性。方法：采用建立的实时荧光定量PCR、常规PCR及传统细菌培养法3种方法,同时对单核细胞增生性李斯特菌等细菌进行检测。结果：实时荧光定量PCR检测的灵敏度可达19 cfu/ml,且有很高的特异性,对英诺克李斯特菌等10种相关细菌均无交叉反应,从细菌核酸提取至完成检测仅需3 h左右。结论：实时荧光定量PCR检测由于在密封环境中进行,避免了产物与环境间的交叉污染,且是3种方法中最为快速敏感的方法,适用于公共卫生应急疫情的实验室快速诊断。

Comparison of real-time fluorescence PCR with PCR and bacterium culture in detection of Listeia monocytogenes

Objective:To compare the sensitivity and specitivity of real-time PCR and PCR and bacterium culture for Listeia monocytogenes detection.Methods:A one-tube real-time fluorescence PCR with the ABI7300 for Listeia monocytogenes detection,PCR and Bacterium culture were performed.Results:The specificity of the assay was high and there were no cross reactions with Listeia innocua.The sensitivity of the assay was 19 cfu/ml and bacterium DNA was directly detected from the clinical specimens by this assay.It took only three hours to extract bacterium DNA and do the real-time PCR.Conclusion:The high sensitivity and specificity and the rapid turnaround time made real-time PCR valuable for the rapid diagnosis of Listeia monocytogenes,especially in a public health laboratory.The closed real-time PCR system avoids possible cross-contamination with PCR and the excessive manipulations required for conventional PCR analysis and saves time and labor.