Development and prevalidation of two different ELISA systems for the potency testing of Clostridium perfringens β- and ϵ-toxoid containing veterinary vaccines

The potency testing of Clostridium perfringens mono- and multicomponent veterinary vaccines is currently performed with the mouse neutralisation test (MNT) to estimate levels of C. perfringens β- and ϵ-antitoxin levels in the sera of rabbits immunised with the vaccine. Two in vitro methods based on monoclonal antibodies (mAb) have been developed for the determination of specific antibodies against C. perfringens β-toxin (capture ELISA) and ϵ-toxin (competitive ELISA) in these sera. Both test systems show high specificity and good reproducibility. These ELISA procedures were used in addition to the routine batch potency test in mice (MNT) to determine β- and ϵ-antitoxin levels in 523 samples of rabbit serum. There was good agreement between the rank order of sera determined in vivo and the rank order determined in vitro. Linear regression analysis gave correlation coefficients of 0.88 for the capture ELISA and 0.41 for the competitive ELISA, with a significance level of P