检测牛奶中肠出血性大肠杆菌O157:H7的双抗夹心化学酶联免疫方法的建立  

Development of a double-antibody sandwich chemiluminescence enzyme immunoassay for detection of Escherichia coli O157:H7 in milk

作　　者：辛思培 蒋蔚[2] 龙梦瑶 李思 王权[2] 孙卫东[1] XIN Sipei;JIANG Wei;LONG Mengyao;LI Si;WANG Quan;SUN Weidong(College of Veterinary Medieine of Nanjing Agricultural University,Nanjing 210095,China;Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《畜牧与兽医》2020年第4期116-121,共6页Animal Husbandry & Veterinary Medicine

基　　金：“十三五”国家重点研发计划项目子课题(2016YFD0501101)。

摘　　要：为了检测牛奶中肠出血性大肠杆菌(enterohemrrhagic Escherichia coil, EHEC)O157:H7污染,采用兔多抗作为捕捉抗体与一株O157:H7单克隆抗体作为检测抗体进行配对,构建了针对EHEC O157:H7的双抗夹心化学发光酶联免疫检测方法。该方法最低检测限可达到2.5×10~4 CFU/mL,优于ELISA检测方法,且特异性良好,与其他大肠杆菌、沙门菌、李斯特菌、阪崎肠杆菌等均无交叉反应。在用于牛奶样本的模拟添加检测中,较ELISA方法可缩短前增菌处理时间。本研究为牛奶中O157:H7污染检测提供了一种准确度高、特异性强的检测手段。In order to detect Escherichia coli O157:H7 contamination in milk, rabbit polyclonal antibodies were used as capturing antibodies, and a monoclonal antibody against E. coli O157:H7 was used as a detecting antibody. A double antibody sandwich chemiluminescent enzyme immunoassay was constructed for detection of E. coli O157:H7 using the above two antibodies. The results indicated a good specificity of this method with a better detection limit than ELISA, of up to 2.5×10~4CFU/mL. In addition, this method produced no cross reaction with other E. coli, Salmonella, Listeria, E. sakazakii, and so on. In detecting E. coli O157:H7 in the milk samples, the present method required shorter pretreatment time than ELISA did. In conclusion, this study provided a highly accurate and specific method for detection of E. coli O157:H7 in milk.

关 键 词：大肠杆菌O157:H7 单克隆抗体 化学发光酶联免疫法 

分 类 号：S852[农业科学—基础兽医学]