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| Preparation and Establishment of ELISA Method for Detection of Edwardsiella tarda eseB Protein Antibody |
| LI Jun-wei,GAO Fan,WANG Bin |
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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 .
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Received: 25 June 2014
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