Comparison of Indirect ELISA and MNT method for detection and evaluation of Clostridium perfringens epsilon antitoxin in vaccinated rabbits

Introduction and Objectives: Clostridium perfringens is a spore forming anaerobic bacterium which divided into five types based on the production of major toxins. Epsilon toxin is one of the major toxins which are produced by C. perfringens B and D types. Epsilon toxin is the responsible for enteric diseases, generically called enterotoxaemia, in sheep, goats, and other animals. Therefore, evaluation of epsilon antitoxin is urgent for diagnosis and evaluating of the vaccine. MNT has some problem include large number of used animals and Long-duration testing. The purpose of this study was to evaluate the ELISA as an alternative method for measuring of epsilon antitoxin.Materials and Methods: First, epsilon toxin was purified and amount of protein was measured using Lowry method. Then, positive and negative control antisera and samples antisera were prepared. Finally, we were measured epsilon antitoxin by ELISA and MNT simultaneously and the results were analyzed by SPSS software. Results: The negative control cut-off was estimated 0.485 and cross-examination test was shown that epsilon toxin had not cross reaction with other toxins of Clostridium spp. The results of this study showed that there is a significant agreement between In vivo and In vitro tests. Linear regression analysis gave significant correlation coefficients of 0.697 for the indirect ELISA (P<0.01). Conclusion: Finally, ELISA assay could be used as an alternative MNT test. However, further research for evaluation of epsilon antitoxin of Clostridium perfringens is needed for target animals.

کلیدواژه ها:

ELISA ، enterotoxaemia ، Clostridium perfringens ، epsilon antitoxin ، MNT