四种快速检测大肠杆菌O_(157):H_7的方法研究

Comparison of Four Metods in Rapid detection of Enterohemorrhagic Escherichia coli O_(157):H_7 strain

目的:将磁免疫分离技术(immunomagnetic separation,IMS)与显色培养(chromogenic media,Ch)和荧光定量(Re-al-time,R-t)PCR技术结合建立快速检测肉制品中EHECO157:H7的方法。方法:应用四种快速检测方法检测模拟样品和423份肉制品中的EHECO157:H7,比较不同方法的灵敏度和特异度。结果:IMS-Ch法特异性最好,24 h对目标菌含量1-5 CFU/10 g的样品检出率为95%。IMS-PCR对实际样品检测效果评价的尤等指数最高。结论:IMS-PCR是一种可信的快速筛选方法,IMS-Ch法为灵敏、快速的确证方法。

Objective：To develop a method for the rapid detection of EHECO157：H7,Immunomagnetic separation technique were carried out to capture pathogens and extract DNA,this assay combined with fluorescence PCR and chromogenic media technique.Methods： Four methods in rapid detection were used to detcect EHECO157：H7 strain in meat products,the sensitivity and specificity was evaluated and compared.Results： It was demonstrated that IMS-Ch assay was proved to be 100% specific.When the bacteria concentration was as low as 1-5 CFU/10 g in simulated samples,95% positive rate of EHECO157：H7 was obtained by IMS-Ch assay.However,the Youden′s Index of IMS-PCR assay 95%.Conclusion： It is evident that IMS-PCR assay is a new approach for the rapid and correct detection in food,and IMS-Ch assay is a more sensitive and rapid confimatinal method.