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中文摘要：
为快速检测鸭疫里氏杆菌，以纯化的兔抗1型鸭疫里氏杆菌抗体作为捕获抗体，以纯化的鸭疫里氏杆菌单克隆抗体作为检测抗体，建立了双抗体夹心ELISA方法。最佳的方案为用184ng／孔纯化的多抗包被过夜，100mL／L小牛血清37℃封闭2h，检测抗原、单抗、酶标抗体均为37℃作用1h，显色20min后终止反应，D450nm值高于0．330且P／N〉2．1判定为阳性。该方法特异性强，与禽源巴氏杆菌、大肠杆菌、沙门菌无交叉反应，最低检测剂量为10^4CFU／mL。与传统细菌分离方法相比，该方法的敏感性为100％，符合率为85．2％。本研究首次建立了检测鸭疫里氏杆菌的双抗体夹心ELISA方法，且特异、敏感。
英文摘要：
The aim of the present study was to develop a rapid and accurate method for the detection of Riemerella anatipestifer based on a double antibody sandwich ELISA. Purified polyclonal antibody against serotype 1 R. anatipestifer was used as a capture antibody, while purified monoclonal antibody 5G7 was used as a detection antibody. The optimal conditions for the ELISA were as follows：polyclonal antibody with 184 ng per well was added to coat overnight,the plate was blocked by 100 mL/L neonatal calf serum for 2 hours at 37℃ ,the incubation times for detection antigen,monoclonal antibody and second labeled an- tibody were 1 hour at 37℃for each,the value of D450nm was obtained after 20 min coloration. This method has no cross-reaction with poultry-origin Pasteurella multocida, Escherichia coli and Salmonella, and its minimum detectable limit was 104 CFU/mL. The sensitivity of the method was 100% ,and the coincidence between bacterial isolation method and the double antibody sandwich ELISA was 85.2M. Therefore,this study provides a rapid, specific and sensitive detection method for R. anatipestifer.
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