摘要
目的研究不同用量卵磷脂中和剂和PB缓冲液,在双价肾综合征出血热灭活疫苗(Vero细胞)无菌检查中的应用。方法取双价肾综合征出血热灭活疫苗(Vero细胞)转移至卵磷脂中和剂中(摇匀后放置5 min),按薄膜过滤法过滤后,用0.15 mol/L PB缓冲液(30~35℃保温)冲洗滤膜,加入液体培养基培养,进行不同用量卵磷脂中和剂和PB缓冲液适用性试验。用选定的试验条件进行无菌检查方法的适用性试验(薄膜过滤法)及阳性对照验证试验。结果采用10 ml卵磷脂中和剂中和10瓶供试品(摇匀后放置5 min),用0.15 mol/L PB缓冲液(30~35℃保温)冲洗滤膜,每张滤膜每次冲洗量为100 ml,共冲洗2次(末次冲洗液中含小于100 cfu的菌液),该方法过滤速度快,卵磷脂冲洗完全,无残留、不影响结果判定,确定适用。无菌检查方法的适用性试验中,供试品各滤筒中的试验菌与阳性对照比较均生长良好,达到《中国药典》三部(2015版)要求,应用于双价肾综合征出血热灭活疫苗(Vero细胞)的无菌检查中无抑菌作用或其抑菌作用可忽略不计。以金黄葡萄球菌和黑曲霉作为阳性对照菌,0和17 d培养到期后加入阳性菌,对比试验显示试验菌均生长良好。结论该方法适用于双价肾综合征出血热灭活疫苗(Vero细胞)无菌检查。
Objective To investigate the application of different dosages of lecithin neutralizer and PB buffer in the sterility test on bivalent inactivated kidney syndrome hemorrhagic fever vaccine（Vero cells）. Methods Bivalent inactivated kidney syndrome hemorrhagic fever vaccine（Vero cells） was transferred to different concentrations of lecithin neutralizer, shaken well, placed for 5 min and filtered by membrane filtration method. The filter membrane were rinsed with 0. 15 mol/L PB buffer（30 ~ 35 ℃）, cultured in liquid medium, and subjected to suitability test on different dosages of lecithin neutralizer and PB buffer. The suitability test on method for sterility test（membrane filter method）and positive control verification test were performed under the selected condition. Results Ten containers of test samples which were shaken well and placed for 5 min were neutralized with 10 ml lecithin neutralizer. Each piece of filter membrane was rinsed with 100 ml of 0. 15 mol/L PB buffer（30 ~ 35 ℃） for 2 times（the bacterial content in the last flushing was less than 100 cfu）. The method was rapid in filtration, while the lecithin was rinsed completely without residue, which showed no influence on the judgment of result. In the suitability test on method for sterility test, the test bacteria in various filter cartridges grew well as compared with positive control, which met the requirements in Chinese Pharmacopoeia（Volume Ⅲ, 2015 edition）, indicating no or negligible bacteriostatic effect. In the positive control verification test by addition of Staphylococcus aureus and Aspergillus niger on days 0 and 17, the test bacteria grew well.Conclusion The method was suitable for the sterility test on bivalent inactivated kidney syndrome hemorrhagic fever vaccines（Vero cells）.
出处
《中国生物制品学杂志》
CAS
CSCD
2017年第12期1335-1339,共5页
Chinese Journal of Biologicals
