摘要以重组的eseB蛋白为抗原，通过免疫新西兰兔获得了高效价的免疫血清，并建立快速检测迟钝爱德华氏菌的间接ELISA方法。结果显示，用重组eseB蛋白免疫新西兰兔后，ELISA检测血清效价为1∶512000；Western blot检测重组eseB蛋白与多克隆抗体有良好的特异性；通过ELISA检测确定迟钝爱德华氏菌抗原的最佳包被密度为108个／m L ，多克隆抗血清的最佳稀释度为1∶16000，确定其检测迟钝爱德华氏菌的最佳灵敏度为104个／m L ；对不同来源迟钝爱德华氏菌eseB蛋白进行检测，弱毒株15947和ET V与抗血清反应结果为阴性，提示eseB蛋白的表达与其致病性相关，该方法具有较高的特异性。
Abstract：In this study ,high titer of antiserum was obtained by immunizing New Zealand rabbits using the recombinant eseB protein as an antigen ,and indirect ELISA method was established to quickly detect Edwardsiella tarda .It was found that the ELISA titer of the rabbit serum was 1 ∶ 51200 ,and the recombinant eseB protein was shown to specifically recognize the polyclonal antibody by western blotting . The optimal coating concentration of the pathogenic bacterium tested by ELISA assay was found 108 cell/mL ,and the best dilution of polyclonal anti‐serum was 1 ∶ 16000 .The minimum concentration of the pathogenic bacterium was 104 cell/mL ,and different sources of eseB protein of E .tarda were detected , showing that the low virulent strains 15 947 and ETV reacted negatively with the anti‐serum ,indicating that the expression of eseB protein was associated with its pathogenicity .The ELISA method has strong specificity in detecting Edw ardsiella tarda .
收稿日期: 2014-06-25
引用本文:
李军伟,高璠,王斌. 迟钝爱德华氏菌 eseB 抗体制备及免疫检测方法的建立[J]. 水产科学, 2014, (6): 369-373.
LI Jun-wei,GAO Fan,WANG Bin. Preparation and Establishment of ELISA Method for Detection of Edwardsiella tarda eseB Protein Antibody. Fisheries Science, 2014, (6): 369-373.
