胶体金免疫层析法联检食品中5种典型沙门氏菌模型的建立和优化

Preparation and Optimization of Colloidal Gold Strip for Simultaneously Detecting Five Typical Salmonella in Foods

采用胶体金标记抗沙门氏菌单克隆抗体,将沙门氏菌单克隆抗体和驴抗鼠抗体(二抗)喷涂于硝酸纤维膜上分别作为检测线和质控线,研制能联检5种典型沙门氏菌(甲型副伤寒沙门氏菌、鼠伤寒沙门氏菌、猪霍乱沙门氏菌、肠炎沙门氏菌、鸭沙门氏菌)的胶体金免疫层析试纸条。结果表明,该试纸条能达到同时检测5种沙门氏菌的目的,其中甲型副伤寒沙门氏菌的检测灵敏度最高,为105 CFU/m L;鼠伤寒沙门氏菌、猪霍乱沙门氏菌、肠炎沙门氏菌和鸭沙门氏菌的检测灵敏度为106 CFU/m L。沙门氏菌加入量为10~100 CFU时,增菌16~20 h,该试纸条能够检测出牛奶、鸡蛋样本中的5种沙门氏菌。本实验研制的试纸条能快速高效地联检食品中5种典型的沙门氏菌,特异性高、灵敏度好,在对多种沙门氏菌进行联检方面有很重要的应用价值。

Colloidal gold immunochromatographic assay was chosen to simultaneously detect five typical Salmonella(Salmonella paratyphi A, Salmonella typhimurium, Salmonella choleraesuis, Salmonella enteritidis and Salmonella anatum) in foods. In this assay, anti-Salmonella monoclonal antibody was labeled with colloidal gold while anti-Salmonella monoclonal antibody and donkey anti-mouse Ig G were dispensed onto nitrocellulose membrane as the test line and the control line, respectively. The sensitivity of the developed strip to Salmonella paratyphi A was 105 CFU/m L and to the other four strains was 106 CFU/m L. When food samples(e.g., milk and egg) were inoculated with 10–100 CFU Salmonella and cultured for 16–20 h, positive results appeared on the test line of the strip. The developed method is rapid, efficient, sensitive and specific. Therefore, it provides a valuable tool for simultaneously detecting different Salmonella species.