Evaluation of Epsilon Antitoxin of Clostridium Perfringens Type D in The Rabbit Serum By Indirect ELISA

Clostridium perfringens is a gram-positive bacterium divided into five major A, E, and iota toxins based on the four main toxins. Epsilon toxin is a potent clostridial toxin and a major toxin of Clostridium Perfringens B and D types, and is the cause of gangrene gas in humans and enterotoxemia in human. For safety assessment of the vaccine, which is important for prevention of this disease, has been proposed SN (Serum neutralization test) method in the pharmacopoeia. The purpose of this study was to evaluate the ELISA as an alternative method for measuring of epsilon antitoxin. First, vaccine potency test was performed using conventional method and the sera obtained were measured using both conventional and ELISA methods. The negative control cut-off was calculated 0.42 and cross-examination was shown that epsilon toxin had no cross reaction with other toxins of Clostridium spp.The results of this study showed that there is a significant agreement between two tests for serum samples of vaccinated rabbits by polyvalent vaccine. Linear regression analysis gave correlation coefficients of 0.697 for the indirect ELISA, with a significance level of P<0.01. Finally, ELISA test could be used as an alternative MNT test which used for detection antibody in laboratory animals. However, further research in this field is needed for target animals.