餐饮食品中6病原菌叠氮丙啶-定量聚合酶链反应检测方法开发与检验数据分析

Development and data analysis of propidium monoazide-quantitative polymerase chain reaction for detection of 6 pathogenic bacteria in catering food

中文关键词:  叠氮丙啶-定量聚合酶链反应  餐饮食品  沙门氏菌  金黄色葡萄球菌  副溶血性弧菌  单核细胞增生李斯特氏菌  蜡样芽胞杆菌  大肠埃希氏菌O157:H7

英文关键词:propidium monoazide-quantitative polymerase chain reaction  catering food  Salmonella  Staphylococcus aureus  Vibrio parahaemolyticus  Listeria monocytogenes  Bacillus cereus  Escherichia coli O157:H7

中文摘要:

      目的 建立叠氮丙啶-定量聚合酶链反应法(propidium monoazide-quantitative polymerase chain reaction, PMA-qPCR)快速检测餐饮食品中6病原菌的方法。方法 开发6种病原菌前增菌通用型培养基, 采用热裂解方法提取样品DNA, 采用PMA技术鉴别活菌与死菌, 运用分子生物学技术检测样品中目标菌的特异性基因片段, 建立病原菌的PMA-qPCR检测方法, 开展餐饮食品样品病原菌筛查检测。结果 本方法特异性良好, 灵敏度较高, 1533份餐饮食品样品中检出病原菌71株, 检出率为4.63% (71/1533)。结论 本研究运用PMA-qPCR开展病原菌活菌检测技术研究, 所建立研究方法可广泛应用于餐饮食品及相关食品中病原菌的快速筛查检测, 具有较好的应用前景和现实意义。

英文摘要:

      Objective To establish the method for rapid detection of 6 pathogenic bacteria in catering products by propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR). Methods Six kinds of common culture media for preproliferation of pathogenic bacteria were developed, sample DNA was extracted by thermal lysis method, and PMA technology was used to identify living and dead bacteria. Molecular biology technology was used to detect specific gene fragments of target bacteria in the samples. A PMA-qPCR method for detection of pathogenic bacteria was established to carry out pathogen screening and detection of catering food samples. Results This method had good specificity and high sensitivity, and 71 strains of pathogenic bacteria were detected in 1533 catering food samples, with a detection rate of 4.63% (71/1533). Conclusion The established method can be widely used in the rapid screening and detection of pathogenic bacteria in catering food and related food and has a good application prospect.