摘要
选用硝酸纤维素膜作固相载体,建立检测迟钝爱德华氏菌的斑点酶联免疫吸附试验诊断方法。根据棋盘试验,确定迟钝爱德华氏菌免疫血清最佳工作浓度为1∶800,酶标抗体最佳工作浓度为1∶200,以出现明显清晰斑点者判定为阳性。用该方法检测时,迟钝爱德华氏菌呈阳性,温和气单胞菌、嗜水气单胞菌、河流弧菌、溶藻弧菌、鳗弧菌、腐败希瓦氏菌、产碱普罗威斯登菌、阪崎肠杆菌和大肠杆菌均呈阴性。试验结果表明,Dot-ELISA方法简便、特异、快速、结果直观,便于在基层推广使用。
A Dot-Enzyme Linked Immunosorbent Assay(Dot-ELISA)was successfully established for detection of Edwardsiella tarda found in Japanese flounder(Paralichthys olivaceus)by using nitrocellulose membrane as solid carrier.The working concentration of serum samples was 1∶800 dilution and that of the enzyme-labeled goat anti-rabbit IgG was 1∶200 dilution by chessboard assay.The samples with clear and obvious dots were judged as positive reaction.Edwardsiella tarda was positive in the Dot-ELISA,while Aeromonas sobria,A.hydrophila,Vibrio fluvialis,V.alginolyticus,V.anguillarum,Shewanella putrefaciens,Providencia alcalifaciens,Enterobacter sakazakii and Escherichia coli were negative in the assay.The trial indicated that this method was simple,specific,quick,direct,easy extention and application at basic units.
出处
《水产科学》
CAS
北大核心
2010年第3期162-165,共4页
Fisheries Science
基金
唐山市科技局科技攻关项目(06125401A-3)
关键词
斑点酶联免疫吸附试验
牙鲆
迟钝爱德华氏菌
快速检测
Dot-ELISA
Japanese flounder(Paralichthys olivaceus)
Edwardsiella tarda
rapid detection