重组细胞高产型H3N2猪流感病毒株的拯救  

Generation of Cell Culture High-yield Recombinant H3N2 Subtype Swine Influenza Vaccine Candidate by Reverse Genetics

作　　者：杨涛[1] 刘明[1] 刘春国[1] 张云[1] 刘大飞[1] 陈浩[1] 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室农业部动物流感重点开放实验室,哈尔滨150001

出　　处：《病毒学报》2007年第6期471-476,共6页Chinese Journal of Virology

基　　金：国家"973"计划资助项目(2005CB523200);国家"十一五"科技支撑计划资助项目(2006BAD06A05)

摘　　要：为拯救出一株能够在动物传代细胞中高水平复制的H3N2亚型猪流感疫苗株,利用反向遗传操作技术,将A/Goose/Dalian/3/01(H9N2)毒株的PB1、PA、NP、M、NS基因和A/PR/8/34毒株的PB2基因作为内部基因与猪流感病毒A/Swine/Henan/S4/01(H3N2)毒株的HA、NA基因进行重组,成功拯救出了具有高度细胞适应性毒株rH3N2株,该毒株接毒MDCK细胞60h后,血凝价可以达到1∶512,表明该毒株具有高度适应细胞繁殖特性,为H3N2亚型猪流感病毒细胞培养型疫苗的研制奠定了基础。High-yield H3N2 subtype swine influenza virus for large-scale vaccine production in cell culture was generated by reverse genetics. The rescued H3N2 （rH3N2） candidate virus contained hemagglutinin （HA） and neuraminidase （NA） genes derived from a field isolate A/Swine/Henan/S4/01（H3N2）, PB2 gene from A/PR/8/34, and the other five internal genes from A/Goose/Dalian/3/01（H9N2）. The rH3N2 virus titer in MDCK cell culture were measured by hemagglutination assay and the maximum virus titre of 1 ： 512 hemagglutination unit was obtained after infection of MDCK cell for 60 h. The results of the present study indicated that rH3N2 virus was suitable for growth in MDCK cell culture and is leasable to be used for the production of cell grown influenza vaccine.

关 键 词：猪流感 H3N2亚型 反向遗传操作技术 细胞培养型流感疫苗 

分 类 号：S858.28[农业科学—临床兽医学]