无细胞百日咳攻击菌的制备及液氮保存方法分析

为了制备无细胞百日咳效价测定用攻击菌并用液氮进行保存,对此方法进行评价。采用原代攻击菌复苏传代后测定细菌浓度,并用蛋白胨水稀释为27亿个菌/m l,分装冻存管中。放入液氮罐内,并对液氮保存的攻击菌进行相应的检测,用统计软件计算。结果显示,此方法制备的攻击菌与传统的攻击菌无显著性差异,冻后虽攻击菌的毒力有所下降,但都在100-1000/MLD的正常范围内,且冻后0、3、6、9、12月无显著性差异。同批支间差异小,CV<5%。通过此方法制备的攻击菌稳定性好,试验结果符合《中华人民共和国药典》要求,可用于无细胞百日咳的效价测定。

Preparation of acellular pertussis challenge strain and analysis of preservation in liquid nitrogen

To prepare a challenge strain used for evaluation of potency of acellular pertussis vaccine and to preserve it in liquid nitrogen,the original challenge strain was recovered and generated,used in determination concentration of the strain.The challenge strain was diluted to 27 hundred million cells per milliliter and stored in liquid nitrogen and then to detect it by calculation on statistics software.There is no significant difference compared by using the routine method.Although the toxicity of stored stain was reduced in liquid nitrogen,it is still maintained in a normal range of 100 to 1000 MLD(minimum lethal dose).There are no significant difference in the challenge strain after stored for 0,3,6 and 9 months respectively,in addition,variations are lower among vials in the same batch CV<5%.This challenge stain is stable and meets with requirements of "Pharmacopoeia of the People's Republic of China".