鲜猪肉中沙门和金葡菌荧光定量PCR检测  

Real-Time PCR Assay for Salmonella and Staphylococcus Aureus in Pork

作　　者：马凯[1] 白羽[1] 陈尔凝[1] 刘杰[1] 刘悦[1] 武会娟[1] 高丽娟[1] 

机构地区：[1]北京市理化分析测试中心,北京100089

出　　处：《食品研究与开发》2015年第17期117-122,共6页Food Research and Development

基　　金：北京市科技新星(2008B31)

摘　　要：通过结合两种技术-免疫磁分离技术与双重荧光定量PCR技术,建立同时、快速对鲜猪肉进行沙门氏菌和金黄色葡萄球菌的检测方法。利用偶联有特异性抗体免疫磁球,在37℃的条件下能够有效地从250 m L循环体系中边富集边捕获目标菌。利用特异性的引物与探针,对两种食源性致病菌进行双重荧光定量PCR同时检测。本研究方法针对沙门氏菌和金黄色葡萄球菌的检测限分别达到3.6 CFU/g和9.2 CFU/g。方法总体灵敏度、特异性和准确度达到98.8%、100%以及98.9%。本研究建立的免疫磁分离-双重荧光定量PCR方法比国标方法更快速和高效,适用于鲜猪肉中沙门氏菌和金黄色葡萄球菌的快速检测。In this study, a rapid and simultaneous detection for Salmonella sp. and Staphylococcus（S.） aureus from fresh pork samples was developed by combining immunomagnetic separation（IMS） with duplex real-time PCR（RT-PCR）. Magnetic beads coated with specific antibodies were used to capture the pathogens from 250 mL at 37 ℃. Then, duplex RT-PCR was applied with two sets of specific primers and probes. The limit of detections were 3.6 CFU/g for Salmonella, and 9.2 CFU/g for S. aureus. The sensitivity, specificity, and accuracy of IMSmultiplex RT-PCR method were 98.8 %, 100 %, and 98.8 %, respectively. IMS-duplex RT-PCR method is more rapid and effective than GB method, and which was suitable for the rapid detection of Salmonella and Staphylococcus aureus in fresh pork.

关 键 词：沙门氏菌 金黄色葡萄球菌 免疫磁分离 双重荧光定量PCR 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程] O657.3[轻工技术与工程—食品科学与工程]