建立单核细胞增生李斯特菌液相芯片检测方法。本研究将单核细胞增生李斯特菌单克隆抗体（mAb930）与聚苯乙烯微球偶联，结合双抗体夹心技术建立单核细胞增生李斯特菌液相芯片检测方法。测定不同浓度的抗体与微球偶联率。通过L25(56)正交设计试验优化方法的反应条件，并进行灵敏度和特异性试验。应用建立的方法及国家标准检测方法同时检测200份食品样品，比较检测结果。结果显示，较优实验条件即多抗工作浓度为1:100、生物素标记的羊抗兔IgG工作浓度为1:500、SA-PE的工作浓度为10 μg/mL、生物素标记的抗体与SA-PE反应时间为30 min。该方法灵敏度可达103 CFU/mL；与其他常见食源性致病菌无交叉反应。与国家标准方法检测单核细胞增生李斯特菌的结果基本相符，液相芯片法检测各种样品的假阳性率低于0.77%。该方法灵敏度高、特异性强、重复性好，可快速检测食品中的单核细胞增生李斯特菌，能够应用于实际样品的检测。

Listeria monocytogenes liquidchip detection method was established by coupling Listeria monocytogenes monoclonal antibody (mAb930) and polystyrene microspheres, and combining with double antibody sandwich technique. The ratios of different concentration of antibody to the microsphere coupling were detected. L25(56) orthogonal test was used to optimize the reaction conditions, and verification of the sensitivity and specificity were detected by the developed method. 200 food samples were tested by the method, with comparison of the national standard method. The optimal concentrations of polyclonal antibody, , the antibody biotin labeled Goat anti rabbit IgG and SA-PE antibody were 1:100, 1:500 and 10 μg/mL, respectively. And the best reaction time of biotin labeled and SA-PE was, 30 min. The sensitivity of this method reached 103 CFU/mL. No cross reaction with other common food borne pathogenic bacteria occurred. In addition, the results were consistent with the national standard method. The false positive rate of liquid chip assay for the detection of various samples was less than 0.77%. The method has high sensitivity, strong specificity, good reproducibility, and can be used for rapid detection of Listeria monocytogenes in food.

广东出入境检验检疫局科技计划项目（2013GDK02）