Investigation of the use of the ELISA procedure in determining the potency of Clostridium chauvoei vaccines

Authors : Deha Ali Deniz, Şükrü Kırkan

Pages : 883-888

Doi:10.35229/jaes.1812152

View : 47 | Download : 143

Publication Date : 2025-11-30

Article Type : Research Paper

Abstract :Blackleg, caused by Clostridium chauvoei, remains a serious disease of cattle and small ruminants. Potency testing of C. chauvoei vaccines is still based on guinea pig challenge assays, which in this study involved 300 Wistar Albino guinea pigs (350–450 g). Although regarded as the reference method, it is labor-intensive, time-consuming, and raises animal welfare concerns. This study aimed to evaluate whether an antigen-based ELISA could serve as a rapid, reproducible, and animal-friendly in vitro alternative for potency testing of inactivated C. chauvoei vaccines. Twenty commercial vaccines (15 with C. chauvoei antigen, 5 without) were evaluated by both the guinea pig challenge test and a flagellar antigen–specific ELISA. For each vaccine, 10 guinea pigs were vaccinated and 5 remained as controls. Relative potency (RP) was calculated against a standardized reference vaccine. Correlation between methods, sensitivity, and specificity were determined. The ELISA achieved 93.3% sensitivity, 100% specificity, and 95.0% relative accuracy. RP values of positive vaccines ranged from 1.02 to 5.44, showing a strong correlation with in vivo results (r = 0.882, p < 0.001). Intra-plate (CV < 10%) and inter-plate (CV < 15%) variation were within acceptable limits. No false positives occurred, and vaccines lacking C. chauvoei antigen produced very low RP values. The developed ELISA represents a reliable alternative to animal testing, significantly reducing both animal use and testing duration while maintaining accuracy. Implementation would align with 3R principles (Replacement, Reduction, Refinement) and enhance the efficiency of vaccine quality control protocols.
Keywords : Aşı, Clostridium, ELİSA, Potens, Yanıkara.

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