CAJ | 학술논문

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。
이중조적eseB단백위항원，통과면역신서란토획득료고효개적면역혈청，병건립쾌속검측지둔애덕화씨균적간접ELISA방법。결과현시，용중조eseB단백면역신서란토후，ELISA검측혈청효개위1∶512000；Western blot검측중조eseB단백여다극륭항체유량호적특이성；통과ELISA검측학정지둔애덕화씨균항원적최가포피밀도위108개／m L ，다극륭항혈청적최가희석도위1∶16000，학정기검측지둔애덕화씨균적최가령민도위104개／m L ；대불동래원지둔애덕화씨균eseB단백진행검측，약독주15947화ET V여항혈청반응결과위음성，제시eseB단백적표체여기치병성상관，해방법구유교고적특이성。
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 .