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.
出处
《食品研究与开发》
CAS
北大核心
2015年第17期117-122,共6页
Food Research and Development
基金
北京市科技新星(2008B31)
