论文部分内容阅读
目的建立一种快速检测金黄色葡萄球菌的液相芯片检测方法,并进行初步应用。方法将金黄色葡萄球菌单克隆抗体(mAb930)与聚苯乙烯微球偶联,结合双抗体夹心技术建立金黄色葡萄球菌液相芯片检测方法,通过L(934)正交设计试验优化方法的反应条件;对建立的方法进行灵敏度和特异性验证;应用建立的方法检测200份食品样品,并与国家标准检测方法进行比较。结果所建立的检测方法的最佳反应条件为:多克隆抗体工作浓度为1∶100,生物素标记的二抗工作浓度为1∶1 000,链霉亲和素-藻红蛋白的工作浓度为2μg/ml,生物素标记的二抗与SA-PE的反应时间为40 min;建立的方法检测金黄色葡萄球菌的灵敏度可达103CFU/ml,检测其他常见食源性致病菌无交叉反应;建立的方法与国家标准方法检测200份食品样品的结果基本相符。结论已建立了一种快速检测食品中金黄色葡萄球菌的液相芯片检测方法,能够应用于实际样品的检测。
Objective To establish a rapid detection of Staphylococcus aureus liquid chip detection methods, and for preliminary application. Methods Monoclonal antibody against Staphylococcus aureus (mAb930) was conjugated to polystyrene microspheres. The detection of Staphylococcus aureus liquid chromatography chip with double antibody sandwich technique was established. The reaction was optimized by L (934) orthogonal design Conditions; sensitivity and specificity of the established method validation; application of the established method to detect 200 food samples, and compared with the national standard test methods. Results The optimal reaction conditions were as follows: the concentration of polyclonal antibody was 1: 100, the concentration of biotin labeled secondary antibody was 1: 1 000, the concentration of streptavidin-phycoerythrin was 2μg / ml, biotin-labeled secondary antibody and SA-PE reaction time was 40 min; the established method to detect Staphylococcus aureus sensitivity up to 103CFU / ml, detection of other common foodborne pathogens no cross-reaction; The established method is basically consistent with the results of the national standard method for detecting 200 food samples. Conclusion A liquid-phase microarray detection method for rapid detection of Staphylococcus aureus in food has been established and can be applied to the detection of real samples.