Antibody induction test (AIT) is a promising candidate as a refinement of the troublesome National institutes of Health (NIH) test in the sense of animal welfare 3R approach for determination of potency of inactivated rabies vaccines for veterinary and human use. In this study, we initially try to develop AIT as a suitable alternative to NIH test, to achieve a reduction of test duration and diminish animal suffering by omitting intracerebral CVS infection and measuring humoral immunity upon vaccination. Designs of both multi-dose and single-dose AIT were examined. Biological reference preparation, batch 5 with assigned titer of 10 IU/vial, was taken as both standard and test vaccine. Six consecutive AITs were performed and eight pools of sera in each AIT were tested in triplicate by rapid fluorescent focus inhibition test. We estimated the upper detection limit and calculated test variability for individual dilutions. For multi-dose AIT, we estimated the dose–response function and performed calculations of final test results and statistical validity parameters for both linear and sigmoidal model using CombiStats program. Sigmoidal 4-parameter dose–response model was found optimal. Presented design of multi-dose AIT showed a satisfactory detection limit for testing of inactivated rabies vaccines for both veterinary and human use. However, due to nonconformity of obtained results with statistical validity criteria, we concluded that the presented model of multi-dose AIT was unsuitable for introduction in routine practice. However, we concluded that there was a realistic option for introduction of two versions of single-dose AIT. The first version would be with two standard vaccine controls and could be introduced immediately, while the second version would include testing of the sample only and rely on comparison of the induced rabies antibody level with absolute cut-off limits set in advance.

中文翻译：

---

可以立即引入单剂量抗体诱导试验，以完善用于灭活狂犬病疫苗效力测定的NIH试验。

抗体诱导测试（AIT）是对麻烦的美国国立卫生研究院（NIH）测试的改进的有前途的候选者，该测试从动物福利3R方法的意义上确定了兽用和人用灭活狂犬病疫苗的效力。在这项研究中，我们最初尝试开发AIT作为NIH测试的替代方法，以通过减少脑内CVS感染并在接种疫苗后测量体液免疫力来缩短测试时间并减轻动物的痛苦。检查了多剂量和单剂量AIT的设计。将标准滴度为10 IU /瓶的生物参考制剂（第5批）作为标准疫苗和测试疫苗。进行了六个连续的AIT，并通过快速荧光聚焦抑制试验一式三份地测试了每个AIT中的八份血清。我们估算了各个稀释液的检测上限，并计算了测试变异性。对于多剂量AIT，我们估算了剂量反应函数，并使用以下方法对线性和S型模型的最终测试结果和统计有效性参数进行了计算CombiStats程序。乙状结肠四参数剂量反应模型被发现是最佳的。提出的多剂量AIT设计显示出令人满意的检出限，可用于兽用和人用灭活狂犬病疫苗的测试。但是，由于所得结果与统计有效性标准不一致，因此我们得出结论，所提出的多剂量AIT模型不适合在常规实践中引入。但是，我们得出的结论是，引入两个版本的单剂量AIT有一个现实的选择。第一个版本将带有两个标准疫苗对照，并且可以立即引入，而第二个版本将仅包括样品测试，并依赖于诱导狂犬病抗体水平与预先设定的绝对临界值的比较。