PCR检测速冻食品中金黄色葡萄球菌与沙门菌

PCR on detection of Staphylococcus aureus and Salmonella in frozen food

目的探讨多重PCR法在速冻食品中金黄色葡萄球菌与沙门菌检测中的应用效果。方法应用金黄色葡萄球菌ATCC8095与沙门菌ATCC14028对速冻水饺进行人工染菌,分别取不同培养时间点增菌液采用水煮法、酚氯仿法与滤柱法剂盒法提取DNA,分析金黄色葡萄球菌与沙门菌多重PCR法在速冻食品中的检测灵敏度、检出时限及合适的DNA提取方法。结果通过不同持续培养时间检测金黄色葡萄球菌与沙门菌基因组DNA,在持续培养时间超过4 h时应用多重PCR法便可检测出速冻食品中100 CFU/g起始菌落数目的金黄色葡萄球菌与沙门菌。水煮法的产量最高、所需时间最短并且费用最低,而试剂盒法提取纯度最高。结论应用多重PCR法检测速冻食品中黄金色葡萄球菌与沙门菌的效果较好,具备良好的快捷性、高效性及稳定性。

Objective To discuss the effect of multiplex PCR on detection of Staphylococcus aureus and Salmonella in frozen food. Methods The quick frozen dumpling was artificial contaminated by Staphylococcus aureus ATCC8095 and Salmonella ATCC 14028. The DNA was extracted by boiling method, phenol chloroform method and filter column box method in difference incubation time. The detection sensitivity, detection time limit and appropriate DNA extraction method of Staphylococcus aureus and Salmonella multiplex PCR in frozen food were analyzed. Results The genomic DNA of Staphylococcus aureus and Salmonella were detected by different culture time. When the duration of culture was longer than 4 h, in the number of initial colonies of 10^0 CFU/g, Staphylococcus aureus and Salmonella in frozen food were detected by multiplex PCR. The boiling method has the highest yield, the shortest time and the lowest cost, and the highest purity is obtained by the kit method. Conclusion The detect effect of Staphylococcus aureus and Salmonella in frozen food by multiplex PCR is better, it has good quickness, high efficiency and stability.