流感病毒裂解疫苗生产用毒株传代稳定性分析

Stability of influenza virus strain for production of cleavage vaccine during subculture

目的分析流感病毒裂解疫苗生产用H1N1、H3N2和B型流感病毒毒株主种子批、工作种子批及疫苗病毒收获液的传代稳定性。方法对制备的H1N1、H3N2和B型流感病毒毒株主种子批、工作种子批及疫苗病毒收获液进行全面的生物学特性检定;采用RT-PCR法从3个型别流感疫苗生产用毒株主种子批、工作种子批及疫苗病毒收获液中扩增血凝素(hemagglutinin,HA)及神经氨酸酶(neuramidinase,NA)基因片段,进行全基因序列测定分析。结果制备的3个型别流感病毒毒株主种子批、工作种子批和疫苗病毒收获液的抗原性与WHO推荐毒株相一致,生物学特性均符合《中国药典》三部(2010版)标准;HA及NA基因核苷酸序列、氨基酸序列均一致,除H3N2型毒株的NA基因序列外,均与GenBank中登录的相应毒种基因序列同源性为100%。结论本公司流感病毒裂解疫苗毒种制备及疫苗生产工艺,能够保持生产用毒株传代生物学特性的一致性及遗传性状的稳定性。

Objective To analyze the stabilities of seeds from master and working seed lots for production of influenza H1N1, H3N2 and B virus cleavage vaccines during subculture as well as the harvested virus liquid. Methods The seeds from master and working seed lots used for production of influenza H1N1, H3N2 and B virus cleavage vaccines as well as the harvested virus liquid were subjected to overall control tests on biological characters, from which hemagglutinin （HA） and neuramidinase （NA） gene fragments were amplified by RT-PCR and sequenced. Results The antigenicities of seeds from master and working seed lots and the harvested virus liquid were consistent with those of strains recommended by WHO, while the biological characters met the requirements in Chinese Pharmacopoeia （Volume m, 2010 edition）, and the HA and NA gene sequences were in agreement. Except the NA gene sequence of H3N2 virus, all the gene sequence showed the homologies of 100% to those of corresponding virus strains in GenBank. Conclusion The procedures for preparation of seeds and production of vaccine developed by Shanghai Institute of Biological Products Co., Ltd. guaranteed the consistency of biological character and genetic stability of vaccine strains during production.