H1和H3亚型猪流感病毒血凝素单因子血清的制备  

Preparation of hemagglutinin subtype-specific serums against H1 and H3 subtype swine influenza viruses

作　　者：翁少亭 曹建科[1] 常艳凯 张素梅[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]中国农业科学院兰州兽医研究所,甘肃兰州730046

出　　处：《河南农业大学学报》2016年第5期646-650,共5页Journal of Henan Agricultural University

基　　金：河南省科技开放合作项目(132106000065)

摘　　要：为了优化H1和H3亚型猪流感病毒（Swine Influenza Viruse,SIV）血清学检测方法,更准确了解猪流感病毒在中国猪群中的流行与分布情况,本试验制备了欧洲禽源猪H1N1病毒和人源猪H3N2病毒的血凝素特异性单因子血清。将人工合成的2种HA基因与p CAGGS真核表达载体做连接,构建HA基因的真核表达重组质粒。经酶切鉴定及体外瞬时表达检测证实重组质粒结构正确,能正常表达目的蛋白。大量提取纯化重组质粒,以每只鸡200μg的剂量肌肉接种SPF鸡,三免后收获鸡血清。通过血凝抑制试验检测HA单因子血清效价。试验结果显示,HA单因子血清抗体水平达到4log2-6log2。交叉HI试验结果显示,2个亚型血清间无交叉反应现象,特异性强。In order to detect quickly and exactly H1 and H3 subtypes swine influenza viruses by optimizing serological methods,SIV hemagglutinin subtype-specific serums against European avian-like H1 N1 and human-like H3N2 were prepared in this study. The selected HA genes were first synthesized and cloned into eukaryotic expression pCAGGS vector. Then the two recombinant plasmids were confirmed by enzyme digestion identification and indirect immuno fluorescent assay （IFA）. SPF chickens were injected by the recombinant plasmids in a volume of 200 μg and the serum of the chickens was collected after the third immunization. Hemagglutinin inhibition test （HI） indicated that the antibody levels reached 4log2 to 6log2 with no cross-reaction. The subtype-specific serums prepared in this study could be used to identify HI and H3 subtypes SIV and cound provide reference for swine influenza vaccine application in pig herds in China.

关 键 词：猪流感病毒 H1亚型 H3亚型 单因子血清 

分 类 号：S855.3[农业科学—临床兽医学]