人用H5N1禽流感疫苗毒种检定及毒种库建立

目的为了研制人用H5N1禽流感疫苗,按照《中国药典》三部“生物制品生产和检定用菌毒种管理规程”中的要求,建立用于生产禽流感疫苗的毒种库。方法从国外引进2株H5N1禽流感疫苗的毒种（R1194和R1203）,进行全面检定,建立三级种子库,即原代、主代和工作代,用于疫苗生产。结果引进的R1194和R12032个毒株能在胚蛋中大量增殖并稳定传代;与普通流感A1,A3,B作交叉血凝抑制试验和交叉单向免疫扩散试验表明,符合H5N1禽流感病毒的特征;经对血凝素序列测定,证实了引进的2毒株为H5N1禽流感病毒;测得的序列与原始株序列比较可知,2毒株均已去除有毒基因,属弱毒株;确定2毒株的P2为原代,P4为主代,P5为工作代。结论引进的R1194和R1203毒种为H5N1禽流感减毒株,可用于禽流感疫苗的生产。

Study on avian influenza (H5N1) vaccine used in human body and establishment of virus seed bank

Objective To develop avian influenza(H5N1) vaccines used in human body.To establish the virus seed bank used to produce avian influenza vaccines according to "Requirements for Bacterial and Viral Strains-Seeds Used for Production and Quality Control of Biologics" from the book called PHARMACOPOEIA OF THE PEOPLE'S REPUBLIC OF CHINA Volume III.Methods To importe from abroad and checked up entirely the two-virus seeds of R1194 and R1203 for the avian influenza vaccine production.To establish three virus seed lot systems(seed bank) i.e.the primary seed lot,master seed lot and working seed lot.Results The two seeds of avian influenza vaccines can be multiplied greatly and passaged steadily in egg embryos;The cross hemagglutinin-inhibiting test and immunodiffusion test with the antigen and antibody of human influenza A1,A3 and B and avian influenza H5N1 indicated that the above two seeds had the character of avian influenza H5N1.The complete HA genes,that were sequenced,approved that the seeds were avian influenza H5N1 viruses.The virulence genes were deleted and the two seeds belonged to feebleness virulence strain by sequence compared with wild types.The two seeds,which were passed 2,4 and 5 times in egg embryos respectively,were confirmed as the primary seed lot,master seed lot and working seed lot.Conclusion The two-virus strain R1194 and R1203 imported from abroad can be used to produce the avian influenza vaccines.