以迟缓爱德华菌株2CDM001为抗原，利用杂交瘤技术制备了迟缓爱德华菌的单抗8D11、8H4、2F4、18C12、16E2、20D3、19G2、5F7、4E6、7C5、18H9。在特异性方面，单抗8H4和2F4特异性较差，与美人鱼弧菌 ATCC33539、鮰爱德华菌 ATCC33202等参考菌株均出现不同程度的交叉反应，而其余9株单抗特异性强，与实验中用到的除迟缓爱德华菌外的参考菌株均不结合；但与迟缓爱德华菌分离株的检测结果表明，仅有20D3和8H4可与所有分离株结合，其余单抗不能与实验中所有迟缓爱德华菌发生反应。由于单抗8H4与美人鱼弧菌 ATCC33539有交叉反应，因此选单抗20D3和兔源多抗建立迟缓爱德华菌的双抗夹心 ELISA 检测方法，该方法特异性强，灵敏度达到5×107CFU/mL。本研究扩充了迟缓爱德华菌单克隆抗体库，为迟缓爱德华菌单克隆抗体的进一步应用提供更多参考数据和可选择的材料。%Using Edwardsiella tarda 2CDM001 as antigen and hybridoma technique,we prepared the monoclonal antibodies(mAbs) 8D11,8H4,2F4,18C12,16E2,20D3,19G2,5F7,4E6,7C5 and 18H9 against E. tarda. Among these mAbs,8H4 and 2F4 had the poor specificity,presented varied cross reactions with control strains such as Vibrio damsela(ATCC33539)and Vibrio damsela(ATCC33539). The other 9 mAbs had favorable specificity and no cross reactions with control strains except E. tarda used in the experiment. The result of detecting E. tarda isolates showed that only 20D3 and 8H4 reacted with all the E. tarda isolates,and the rest of mAbs did not reacted with all the E. tarda isolates. Due to the cross reaction of 8H4 with V. damsela(ATCC33539),thus 20D3 was selected to establish a double-antibody sandwich ELISA for E. tarda with rabbit polyclonal antibody. The double-antibody sandwich ELISA possessed the strong specificity,and of which the limited detecting concentration reached 5 × 107 CFU /mL. In conclusion,this study expanded the mAbs library for E. tarda,and also provided more reference data and alternative materials for further application of E. tarda mAbs.
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