Designing of an Indirect ELISA Method for the Detection of Beta Antitoxin of Clostridium perfringens Type C in Rabbit Serum

Clostridium perfringens is the causative agent of enterotoxaemia, which is related to severe economic losses. The evaluation of immunity of beta antitoxin, that prevented animals against the disease, is measured using potency (serum neutralization) technique that it has disadvantages. So Enzyme-linked Immunosorbent Assay (ELISA) has been proposed as an alternative to SN. In this research, before and after purification of beta toxin, MLD and amount of protein was measured using Lowry method. Then, positive and negative control antiserum and samples antiserum were prepared. Then, we were measured beta antitoxin by ELISA and serum neutralization simultaneously and the results were analyzed by SPSS software.The negative control cut-off was calculated 0.485 and cross-examination was shown that beta toxin used in this study has no cross reaction with other toxins of Clostridium perfringens and its repeatability was assessed using standard deviation (for Medium specimen: SD=0.033 CV=7.25) The results of this study showed that there is a significant agreement between in vivo and in vitro tests for serum samples of vaccinated rabbits by enterotoxaemia vaccine. Linear regression analysis gave correlation coefficients of 0.84 for the indirect ELISA, with a significance level of p < 0.01. Sensitivity, positive and negative predictive value of ELISA in these samples was 100%, 84.21% and 100%, respectively. Finally, ELISA systems are suitable candidates to replace the MNT used for detection antibody in laboratory animals. However, further research in this field is needed for target animals.