【摘要】 为了检测食品中金黄色葡萄球菌,建立一种基于SYBR GreenⅡ染料的实时荧光定量聚合酶链式反应(qPCR)检测方法。根据金黄色葡萄球菌nuc基因,设计特异引物,优化引物退火温度,检测了特异性和灵敏性,建立了荧光定量PCR检测方法,并利用此方法对肉类和乳制品中的金黄色葡萄球菌进行检测。结果表明:建立qPCR方法具有很好的特异性,其灵敏度可达到3.5×101 CFU/mL,检测到的循环阈值(Cycle threshold,Ct)与金黄色葡萄球菌菌液浓度有良好的线性方程,相关系数R2为0.9997,在肉类和乳制品中金黄色葡萄菌的检测限分别为4.0×101,3.5×101 CFU/mL。试验证明建立的荧光定量PCR检测方法能够快速准确地检测食品中的金黄色葡萄球菌。更多还原

【Abstract】 In order to detect Staphylococcus aureus in food,a real-time fluorescence-based quantitative PCR(qPCR)detection method based on SYBR Green II dye was established.According to the S.aureus nuc gene,the specific primers were designed,The annealing temperature was optimized and the specificity and sensitivity of the qPCR method for detection of S.aureus were investigated.The detection limit of this method was tested in meat and dairy products.Results indicated that with perfect specificity,the detection sensitivity of the qPCR method was 3.5×101 CFU/mL.There was a perfect linear equation between the Ct value and the concentration of S.aureus.The correlation coefficient R2 was 0.9997.The detection limit of S.aureus in meat and dairy products was 4.0×101 CFU/mL and 3.5×101 CFU/mL,respectively.Experiments showed that the qPCR detection method established in this study can be applied to quickly and accurately detect S.aureus in food.更多还原