The most critical parameter for the quality control of the rabies vaccine is potency, which is evaluated by challenge test in mice while using a large animal number. Because the 3Rs concept is applied worldwide, it becomes necessary to develop alternative methods to demonstrate the production consistency of these vaccines and reduce the number of animals used for performing assays. Hence, the present study evaluated the impacts of reducing the number of mice used in the NIH test for such vaccines. A retrospective data analysis compared vaccines tested in the standard test with the results of the reduced test using only the first cages of each dilution and considering the second cages as their replicates. The relevance of the reduced assay was evaluated using Bland- Altman plot and CCC. Reliability was assessed by CV% and confidence intervals, while the impact of the reduced mouse number was evaluated by the analysis of the confidence interval of potency results and regression, linearity and parallelism parameters. The results demonstrated the feasibility of reducing to eight mice per dilution in routine assays, with complete statistical validation of the resulting potency, allowing the number of animals used for the test vaccines to be reduced by 50%.

狂犬病疫苗质量控制的最关键参数是效力，可通过在使用大量动物的情况下对小鼠进行攻击试验来评估其效力。由于3Rs概念已在世界范围内应用，因此有必要开发其他方法来证明这些疫苗的生产一致性并减少用于进行测定的动物数量。因此，本研究评估了减少用于此类疫苗的NIH测试中的小鼠数量的影响。回顾性数据分析将标准测试中测试的疫苗与仅使用每种稀释度的第一个笼子并考虑第二个笼子作为复制品的简化测试结果进行了比较。使用Bland-Altman图和CCC评估简化测定的相关性。通过CV％和置信区间评估可靠性，同时通过分析效价结果和回归，线性和平行度参数的置信区间来评估减少的小鼠数量的影响。结果证明了在常规测定中将每稀释度减少至八只小鼠的可行性，并对所得效力进行了完整的统计验证，从而使用于测试疫苗的动物数量减少了50％。