链球菌病类活疫苗活菌计数参考品的研制

Development of a reference substance for live bacterial count of Streptococcicosis live vaccines

摘要:

研制链球菌病类活疫苗活菌计数参考品，可以更加科学地评价活菌计数结果的准确性和有效性。首先，制备了一批链球菌病类活疫苗活菌计数参考品，对其物理性状、纯粹性、真空度、剩余水分进行检验，并对其均一性、运输稳定性、热稳定性进行测定，另组织3家单位通过协作标定的方式对参考品活菌数进行赋值，用协作标定法统计参考品在12个月内的保存期。参考品的性状检查、纯粹检验、真空度测定和剩余水分测定结果均符合《中国兽药典》的规定；均一性试验结果显示，参考品计数结果的变异系数小于10%，均一性良好；运输稳定性试验证明，参考品在夏季和冬季用泡沫盒加冰袋的方式运输3日内数值仍能保持稳定；加速热稳定性试验验证，参考品在–20 ℃条件下保存3个月、4 ℃条件下保存21 d内均可活菌数稳定；通过协作标定，统计出参考品活菌数的赋值范围为 (8.5–12.1)×107 CFU/支；保存期试验结果证实，参考品在–70 ℃以下保存一年内活菌数可维持稳定状态。链球菌病类活疫苗活菌计数参考品，不仅可以为链球菌病类活疫苗的活菌计数实验提供参照物，而且可以用于评价马丁琼脂培养基的质量，为兽用生物制品质量控制提供保障。

Abstract:

This study attempts to develop a reference substance for the live bacteria count of Streptococcicosis live vaccines in order to evaluate the validity of live bacterial count in inspection and testing. We prepared a batch of live Streptococcus suis reference substance for live bacterial count, tested their physical property, purity, vacuum degree, remaining moisture, and determined their homogeneity, thermal stability and transportation stability. Moreover, we organized collaborative calibration to assign count values to the reference substance and determine the shelf life of the reference substance in 12 months. The results showed that the physical property, the purity, the remaining moisture and the vacuum degree of the reference substance were all in compliance with the requirements of the Chinese Veterinary Pharmacopoeia. The homogeneity test showed that the coefficient of variation of the count of the reference substance was less than 10%, indicating a good homogeneity. Transportation stability test showed that the reference substance remained active after 72 h transportation in summer and winter with the package of styrofoam boxes and ice packs. Thermal stability test showed that the reference substance could be stored for up to 3 months at –20 °C, or up to 21 days at 4 °C. According to the collaborative calibration, the reference vaccine was assigned a count value range of (8.5–12.1)×107 CFU/ampoule. The shelf life test showed that the reference substance was stable for 12 months when stored at –70 °C. The reference substance could provide a reference for the live bacterial count of Streptococcicosis live vaccines. Moreover, it could also be used as a reference to evaluate the quality of corresponding agar media.