Abstract

The major limiting factor in immunodiagnostic development for Fasciolosis detection is the absence of specific, monoclonal or polyclonal, antibodies against Fasciola antigens specifically Fasciola gigantica, which is the main species responsible for Fasciolosis in ruminants in Indonesia. The purpose of this study was to produce polyclonal antibody against excretory/secretory (ES) product of F. gigantica Lombok isolate using rabbit. Four months old rabbits were immunized subcutaneously with 0.5ml (300µg/ml) of the ES product emulsified with 0.5ml Freund’s complete adjuvant and then Freund’s incomplete adjuvant. Serum antibodies were harvested after two-times booster at 1-month interval and determined using enzyme linked-immunosorbent assay (ELISA) and Western blotting. Immunization results evaluated by ELISA show that rabbits can be used as a bioreactor to produce ES antibodies. The anti-ES antibody response could be detected 4 weeks post- immunization, followed by increased humoral response of rabbit after first booster. The optical density (OD450nm) value in the ELISA increased from 0.4 before immunization to 1.184 and 1.392 depending on the type of blocking agent used. The Western blotting results confirm that the ES protein bands were only recognized by rabbit serum samples post-immunization and thus confirmed the ELISA test result.

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