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Development of an indirect ELISA system for diagnosis of porcine edema disease using recombinant modified Stx2e
Jin Hur1, Ho-Kyoung Jung2, Seung-Won Park3
1Department of Veterinary Public Health, College of Veterinary Medicine, Jeonbuk National University, Iksan-si, Jeollabuk-do 54596, Republic of Korea.
Aim:
This study aimed to develop a sensitive and specific recombinant antigen protein indirect enzyme-linked immunosorbent assay (ELISA) kit to detect the Shiga toxin (Stx)-producing Escherichia coli (STEC) antibodies against porcine edema disease (ED).
Methods And Results:
The recombinant antigen was co-expressed with the STEC-derived Stx2e A2-fragment and Stx2e B protein in E. coli BL21(DE3) pLysS cells and purified using maltose-binding protein open columns. We used a Shiga-like toxin 2 antibody to test the specificity of the recombinant antigen in an indirect ELISA, which was detected in antigen-coated wells but not in uncoated wells. We tested the indirect ELISA system using samples from the STEC-immunized pig group, the commercial swine farm group, and healthy aborted fetal pleural effusion group; five and twenty samples, respectively, were positive for STEC in the former, whereas all three samples were negative for STEC in the latter.
Conclusions:
This newly developed indirect ELISA may be a specific method for diagnosing STEC infections in pigs.
