H7N9亚型禽流感病毒反向遗传操作系统的建立及其疫苗候选株的构建  

Establishment of reverse genetics system for H7N9 avian influenza virus and construction of candidate recombinant vaccine strain

作　　者：李媛媛[1] 郭晶[1] 李旭勇[1] 王金良[1] 范俊[1] 梁立滨[1] 邓国华[1] 施建忠[1] 陈化兰[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,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2014年第5期407-409,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金："973"专项(2011CD505000)

摘　　要：为建立H7N9亚型禽流感病毒(AIV)反向遗传操作系统,本研究以H7N9亚型(AIV)A/CK/Shanghai/S1053/2013(CK/53)株为亲本病毒,构建了该病毒株的8质粒反向遗传操作系统,并拯救出救获株rCK/53。全基因组序列测定结果表明,rCK/53与亲本病毒的核苷酸序列完全一致。同时以A/PueaoRico/8/34(PR8)的内部基因为骨架,以CK/53的HA和NA的表面基因为供体,构建H7N9亚型AIV疫苗候选株CK53/PR8,疫苗株的8个基因来源与预期完全一致。对rCK/53以及疫苗候选株CK53/PR8在MDCK和A549两种细胞中进行生物学特性的比较,在A549中复制差异不显著,而在MDCK中48 h和72 h两者复制具有明显差异。rCK/53反向遗传操作系统的建立和疫苗候选株CK53/PR8的构建为进一步开展H7N9亚型AIV跨宿主传播机制、致病机理及进一步的免疫保护实验奠定了基础。To establish the reverse genetics system of the H7N9 avian influenza virus （AIV）, eight plasmid system of A/CK/Shanghai/S1053/2013（CK/53） was constructed and the rCK/53 was successfully rescued, of which the whole genome sequences were identical with the parental virus. In addition, the recombinant vaccine virus of CK53/PR8 was generated which contained HA and NA genes of CK/53 in the back ground of internal genes derived from the vaccine virus strain of A/PuertoRico/8/34 （PR8）. Comparison of the growth of rCK/53 and CK53/PR8 in MDCK and A549 cells showed that there was no significant difference of the viruses in A549 cells, but displayed obvious difference at 48 and 72 hours post-infection in MDCK cells. The results would provide a promising technique support for further study on the pathogenic and crossing host barrier mechanisms of the H7N9 avian influenza virus and on further immune protection experiment.

关 键 词：H7N9 禽流感病毒 反向遗传操作系统 疫苗候选株 

分 类 号：S852.65[农业科学—基础兽医学]