流感病毒裂解疫苗病毒种子批的制备和培养条件的研究

Preparation of strains of split influenza virus vaccine and optimization of condition for culture of influenza virus

目的制备流感病毒裂解疫苗的主代和工作种子批,优化病毒培养条件提高产量,制备高质量的流感病毒裂解疫苗。方法将H1N1、H3N2和B型3株流感病毒分别接种SPF鸡胚,收获尿囊液,制备流感病毒裂解疫苗主代及工作种子批。流感疫苗工作种子批按不同稀释倍数、pH值、收获时间和冷胚时间优化病毒培养条件,制成疫苗,进行产量、安全性和免疫原性研究。结果通过优化不同型别流感病毒的培养条件,提高了抗原的血凝滴度,血凝滴度达到1∶1 024及以上,制备的疫苗具有良好的安全性和免疫原性。结论制备出了高产量和高质量的流感病毒裂解疫苗。

Objective To prepare master strains and work strains of influenza virus.To optimize the conditions for culture of influenza and prepare a high qualitative split influenza virus vaccine.Methods Three strains of influenza virus（H1N1、H3N2 and B）were inoculated into Specific Pathogen Free（SPF）Eggs.The allantoic fluid was harvested for preparing master strains and work strains of influenza virus.The effects of dilution ratio of virus inoculated,cultural time and cooling time for harvest on hemagglutination（HA）titer were evaluated.The split influenza virus vaccine was Produced and tested with＂the Production and Control of Split Influenza Virus Vaccine＂.Results The results showed that the condition for culture of influenza virus was optimized as follows：10-5~10-6 dilution ratio of virus and pH value of 7.8 were inoculated into chick embryo,and harvested in 72 hafter culture by cooling for 16~18 h in 4℃.The titer of influenza virus prepared under the optimal condition was above 1∶1 024.The split influenza virus vaccine met the standard requirements.Conclusion The quality split influenza vaccine with high yield is prepared.