| Standardization of an indirect ELISA test for evaluation carp humoral immune response to Aeromonasis |
| Paper ID : 1042-ICIAQUA |
| Authors |
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Ali Shirazinezhad *1, Fatemeh Dabiri2 1iran,shiraz, sanaye square,Razi Vaccine and Serum Research Institute, Education and Extension Organization 2Razi Vaccine and Serum Research Institute, Education and Extension Organization, Shiraz, Iran |
| Abstract |
| Ali Shirazinezhad 1, Fatemeh Dabiri 1, Fatemeh Saemi 1, Zahra Keliddar-Mohammadi 1, Mohammadhossein Hosseini 1 1. Razi Vaccine and Serum Research Institute, Education and Extension Organization, Shiraz, Iran [email protected] Purpose: Serological tests are applied to evaluate humoral immune response (antibodies) in body fluids. Enzyme linked immunosorbent Assay (ELISA) test is the most used test for this purpose. In the present study an indirect ELISA test was standardized to evaluate antibodies against Aeromonas hydrophila in Carp fish vaccinated against this pathogen. Method: To get positive sera against Aeromonas hydrophilla for standardization of indirect ELISA test, BALB-C mice were immunized with Aeromonas hydrophilla bacterin along with complete Freund's Adjuvant (CFA) at zero day by subcutaneous (SC) injection and the same was repeated at fourteen-day post primary injection (Booster dose) with incomplete Freund's adjuvant. Seven-day post booster dose blood was collected from immunized and un immunized mice, and serum samples were used as through positive and through negative respectively for ELISA tests standardization. ELISA test was standardized using checkerboard method by applying various amount of Antigen (Aeromonas hydrophila bacterin).Various dilution of through positive and through negative pooled sera, as well as different dilution of conjugated antibody, and various color developing times. Results: standardized ELISA is working with 1×10 7 bacteria per well, 1% BSA as blocking, 1:100 serum dilution,1:4000 conjugated antibody dilution and 10 minutes developing time. Conclusions: This locally standardized ELISA test was used to evaluate antibody titer in carp immunized against Aeromonosis. |
| Keywords |
| ELISA, Aeromonas hydrophila, Carp fish |
| Status: Abstract Accepted (Oral Presentation) |