免疫磁珠富集-实时荧光PCR技术在食源性致病菌监测中的应用研究

Immunomagnetic enrichment real-time fluorescent PCR technology in the application of food borne pathogenic bacteria monitoring

目的建立单核细胞增生李斯特菌、金黄色葡萄球菌、沙门菌3种致病菌的免疫磁珠富集-实时荧PCR(IMSqPCR)技术的检测方法,应用于食品微生物及其致病因子的监测工作中。方法将菌株及样品稀释后,使用IMSqPCR技术进行检测,用菌株对该方法的特异性进行验证;对586份各类食品用IMS-qPCR和传统培养法进行比较分析。结果 IMS-qPCR方法对13株标准菌株符合率为100.00%;与分离到的3种致病菌的符合率均为100.00%;586份监测食品的检测结果显示,IMS-qPCR方法对食品中3种食源性致病菌的符合率为93.22%,而IMS-qPCR法可在2~3 h内获得检测结果。结论 IMS-qPCR方法可以推广应用于食品微生物及其致病因子的监测,该方法具有特异性强、敏感度高、速度快的特点,也可以应用于突发公共卫生事件应急检测,为事件的处置提供有效技术支撑。

Objective Establishment of Listeria monocytogenes, Staphylococcus aureus, Salmonella three induced detection method of pathogen IMS- qPCR technology application in the monitoring of food microorganism and pathogenic factor. Methods After dilution of the strains and samples,the IMS- qPCR technology was used to test the specificity of the method. 2014- 2015 years of food microbiology and pathogenic factors of the monitoring work,the 586 kinds of food with IMS- qPCR and traditional culture method for comparative analysis. Results IMS- qPCR method of 13 strains of standard with rate was 100. 00%; and work were isolated from the 3 kinds of pathogenic bacteria of the coincidence rate was 100. 00%; 586 food monitoring test results show that,IMS- qPCR method of food in three species of food borne pathogens coincidence rate was 93. 22%,and IMS- qPCR can in 2 ～ 3 h in test results are obtained. Conclusion IMS- qPCR method can be applied to the monitoring of food microorganism and pathogenic factor; the method has strong specificity and high sensitivity,speed can also be applied to detection of public health emergency response,provide effective technical support for the disposal of the incident.