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

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

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。

Objective ：To compare the sensitivity and spocitivity 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 speci- mens 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.