摘要
目的建立一种能同时检测食品中沙门氏菌、大肠杆菌O157∶H7、金黄色葡萄球菌、单核细胞增生李斯特菌和蜡样芽胞杆菌的多重PCR快速检测方法。方法分别针对沙门氏菌侵袭基因invA、大肠杆菌O157∶H7肠溶血素A基因HlyA、金黄色葡萄球菌耐热性核酸酶基因nuc、单核细胞增生李斯特菌李氏溶血素O基因hlyA和蜡样芽孢杆菌肠毒素FM基因entFM序列设计引物,进行PCR扩增及电泳检测。同时优化反应体系,测定特异性和灵敏性。结果初步建立的多重PCR方法可简便、快速、灵敏地实现对5种致病菌的同时检测,整个检测过程少于30h,且检测灵敏度均可达102CFU/ml。结论这种方法是对传统检测方法的有效改进,为食源性致病菌的快速高通量检测提供了理想手段,有良好的应用前景。
Objective A multiple PCR method for rapid and sensitive detection was constructed,which can detcet salmonella,Escherichia coli O157∶H7,Staphylococcus aureus,Listeriosis monocytogenes and Bacillus cereus simultaneously. Method This multiple PCR method was used for primers design,and PCR amplification and electrophoresis,which was aimed at salmonella invasion gene invA,Escherichia coli O157∶H7 intestine hemolysin gene A HlyA,Staphylococcus aureus thermostable nuclease gene nuc, Listeriosis monocytogenes hemolysin O gene hlyA and Bacillus cereus enterotoxin FM gene entFM. Besides, this method can optimize the reaction system and detect the specificity and sensitivity. Results The results indicated that with the convenient, rapid and sensitive method, the detection of the five pathogenic bacteria can be performed together in 30 h. Besides, the sensitivity can reach 102 CFU/ml. Conclusion The approach improves the traditional detecting methods, provides ideal means for the rapid and high-throughput detection of foodborn pathogenic bacteria, and has favourable application perspective.
出处
《中国食品卫生杂志》
北大核心
2009年第5期398-402,共5页
Chinese Journal of Food Hygiene
基金
黑龙江省卫生厅计划项目(2002-107)
黑龙江省科技攻关项目(GC04C42501)
关键词
食源性致病菌
聚合酶链反应
快速检测
Food-borne Pathogens
Polymerase Chain Reaction
Rapid Detection
