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针对食品中金黄色葡萄球菌检测,选取金黄色葡萄球菌耐热核酸酶nuc基因作为靶标,利用地高辛标记引物扩增的PCR产物与生物素标记的探针杂交,然后与ELISA平板上的链霉素杂交,通过抗地高辛抗体显色建立PCR-ELISA检测技术。应用该方法检测人工污染牛奶中金黄色葡萄球菌,同时将大肠埃希氏菌、沙门氏菌、志贺氏菌、单增李斯特菌等常见食源性致病菌作为阴性对照。结果显示,金黄色葡萄球菌的纯培养物以及人工污染金黄色葡萄球菌牛奶,金黄色葡萄球菌为阳性,其他对照菌为阴性,两者PCR-ELISA检测敏感性均为10~1CFU/m L,比普通PCR检测的敏感性(10~3CFU/m L)高100倍。因此,该研究建立的PCR-ELISA方法具有良好的特异性与敏感性,能够特异、敏感、准确地检测牛奶中的金黄色葡萄球菌,为食品中金黄色葡萄菌的快速鉴定、风险评估与有效监测提供重要技术手段。
For the detection of Staphylococcus aureus in food, the nuc gene of Staphylococcus aureus heat-resistant nuclease was selected as the target, the PCR product amplified by digoxin-labeled primer was hybridized with biotin-labeled probe, Mycotoxin hybridization, by anti-digoxin antibody color developed by PCR-ELISA detection technology. The method was applied to detect Staphylococcus aureus in artificial contaminated milk, and common food-borne pathogens such as Escherichia coli, Salmonella, Shigella and Listeria monocytogenes were used as negative control. The results showed that the pure cultures of Staphylococcus aureus, as well as the staphylococcus aureus milk and staphylococcus aureus were positive, while the other control bacteria were negative. The sensitivity of PCR-ELISA and ELISA were both 10 ~ 1CFU / m L, 100 times higher than the sensitivity of ordinary PCR detection (10 ~ 3CFU / m L). Therefore, the PCR-ELISA method established in this study has a good specificity and sensitivity, can be specific, sensitive and accurate detection of Staphylococcus aureus in milk for the rapid identification of food Staphylococcus aureus, risk assessment and effective Monitoring provides important technical means.