【摘要】 应用间接酶联免疫吸附试验(Indirect-ELISA)技术,建立了快速检测柱状黄杆菌的方法。结果显示:根据棋盘试验,确定最佳抗原包被浓度和免疫血清最佳工作浓度分别为1×106cfu/mL和1∶5000,酶标抗体最佳工作浓度为1∶10000。在该条件下,所建立的间接ELISA能检测出5×104cfu/mL的柱状黄杆菌,且与爱德华氏菌、哈维氏菌弧菌、嗜水气单胞菌、副溶血弧菌等常见鱼类致病菌无交叉反应。结果表明,所建立的间接ELISA可不经细菌培养而直接检测人工感染后草鱼鳃组织中的柱状黄杆菌,该方法对柱状黄杆菌的检测具有快速、敏感、特异、实用的优点。更多还原

【Abstract】 This study developed an indirect Enzyme-Linked Immunosorbent Assay(Indirect-ELISA) for the rapid detection of Flavobacterium columnaris.The test indicated that the best working dilutions of antigen and antiserum were 1×106 cfu/mL and 1∶5000 separately,which was determined by using checkerboard titration;the best working dilutions of goat anti-rabbit IgG-HRP affinity purified was 1∶10000.In this condition,the detection limit of Indirect-ELISA was approximately 5×104 cfu/mL.Besides,no cross-reaction was observed when Aeromonas hydrophila,Edwarsiella tarda,Vibrioparah aemolyticu,Vibrio harveyi Aeromonas Sobila were applied instead of F.columnaris.Indirect-ELISA can be directly used to detect F.columnaris ingill of grass carp after challenged with F.columnaris.It showed that the Indirec-ELISA is quick,sensitive,specific,effective,and can be used to detect F.columnaris.infection.更多还原