H5亚型禽流感重组鸡痘病毒活载体疫苗的构建及其遗传稳定性与免疫效力  

Construction,Genetic Stability and Protective Efficacy of Recombinant Fowlpox Virus Expressing Hemagglutinin Gene of H5N1 Subtype Avian Influenza Virus

作　　者：贾立军[1] 彭大新[1] 张艳梅[1] 刘红旗[1] 刘秀梵[1] 程坚 陈素娟 韦栋平 黄勇 张如宽 

机构地区：[1]扬州大学畜牧兽医学院,扬州225009

出　　处：《微生物学报》2003年第6期722-727,共6页Acta Microbiologica Sinica

基　　金：国家"8 63计划"(2 0 0 1AA2 1 3 0 41 ) ;农业部农办牧 2 0 0 1 83项目~~

摘　　要：以鹅源H5亚型禽流感病毒 (AIV)基因组为模板 ,用RT PCR扩增血凝素 (Hemaggluti nin ,HA)基因 ,克隆入鸡痘病毒表达载体pFG1 1 75 ,转染鸡痘病毒感染的鸡胚成纤维细胞 ,通过蓝斑筛选和间接免疫荧光检测 ,获得表达HA基因的重组鸡痘病毒 (Recombinantfowlpoxvir us,rFPV HA)。rFPV HA经鸡胚成纤维细胞连续传 1 5代后 ,报告基因LacZ和HA基因可稳定表达。用 1 0 3PFU和 1 0 5PFU的rFPV HA免疫无特定病原体的 (Specificpathogenfree,SPF)鸡 ,免疫后 2 2d血凝抑制 (Hemagglutinininhibition ,HI)抗体监测阳性率分别为 0 %和 2 0 % ,但均抵御了H5亚型毒株的致死性攻击 ,保护率为 1 0 0 %。结果表明 ,构建了表达HA基因的重组鸡痘病毒 ,该重组病毒具有良好遗传稳定性 ,免疫鸡可提供完全保护 ,显示出了一定的应用前景。Construction, genetic stability and protective efficacy of a recombinant fowlpox virus expressing hemagglutinin(HA) gene (rFPV-HA) of H5 subtype avian influenza virus were conducted. The results indicated that the recombinant fowlpox virus was obtained and purified by blue plaque selection. HA expressed by rFPV-HA was identified by indirect immunofluorescence array. The rFPV-HA could express reporter gene LacZ and HA gene after fifteen successive passages in chicken embryo fibroblast. The rFPV-HA induced completely protection of chickens against lethal challenge with homologous avian influenza virus. The results show that rFPV-HA has good genetic stability and antigenicity and should be a useful tool in HPAI control program by preventing illness and death in chickens when the disease spreads widely.

关 键 词：H5亚型 禽流感 血凝素基因 重组鸡痘病毒 遗传稳定性 免疫效力 

分 类 号：S859.797[农业科学—临床兽医学]