H5N6亚型禽流感病毒反向遗传疫苗株的构建及免疫保护试验  

Construction of Reverse Genetic Vaccine Strain against Variant H5N6 Avian Influenza Virus and its Protective Efficacy Test

作　　者：蒋文明[1] 侯广宇[1] 王素春[1] 李金平[1] 刘朔[1] 陈继明[1] 

机构地区：[1]中国动物卫生与流行病学中心,山东青岛266032

出　　处：《中国动物检疫》2015年第1期64-67,共4页China Animal Health Inspection

摘　　要：为应对抗原性已发生变异的第2.3.4.6分支H5N6亚型禽流感病毒(AIV)引发的潜在流行,本研究利用反向遗传操作技术,以A/Puerto Rico/8/34(PR8)的内部基因为骨架,以H5N6亚型AIV A/Chicken/SC/6/2014(C6)的基因组为模板,经RT-PCR扩增其HA及NA基因,并对HA基因进行分子修饰,去除与H5亚型AIV致病力有关的HA蛋白裂解位点处的多个碱性氨基酸,使其获得低致病性AIV的分子特征(即将-PLRERRRKR-突变为-PQRETR-),成功构建了H5N6亚型AIV疫苗候选株r C6。免疫效力试验表明,r C6重组灭活疫苗可以诱导产生高水平的血凝抑制抗体。免疫攻毒保护试验表明,r C6重组灭活疫苗可以提供SPF鸡抵抗同源和异源H5N6病毒100%的保护,而针对第2.3.4分支的Re-5疫苗仅能提供大约10%的保护。利用反向遗传技术构建的r C6重组疫苗候选株为该分支病毒的防控提供了有益的尝试。To control of H5N6 subtype highly pathogenic avian influenza viruses（HPAIVs） with antigenic changes（clade 2.3.4.6）,and the potential pandemic threat to poultry,a reassortant virus,named r C6,was constructed using reverse genetics in the study. The virus contained modified HA and NA genes of an epidemic strain A/Chicken/SC/6/2014 with the internal genes derived from A/Puerto Rico/8/34（PR8）. The reassortant virus was attenuated by removal of the multi-basic amino acid motif in the HA gene by mutation and deletion（from PLRERRRKR to PQRETR）,thus resulting in a candidate vaccine virus. Immune efficacy tests showed that the inactivated r C6 recombinant vaccine could induce high levels of hemagglutination inhibition antibody. Challenge tests showed that r C6 recombinant inactivated vaccines provided SPF chickens with 100% protection against homologous and heterologous H5N6 virus,and Re-5 vaccine against clade 2.3.4 viruses only provided about 10% protection. The r C6 recombinant vaccine candidate strains using reverse genetics technology provided a beneficial attempt for prevention and control of the virus of clade 2.3.4.6.

关 键 词：禽流感病毒 变异株 H5N6 反向遗传 疫苗 

分 类 号：S852.657[农业科学—基础兽医学]