H9亚型禽流感病毒血凝素特异性单因子血清制备  

Preparation of hemagglutinin antiserum against H9 subtype avian influenza virus

作　　者：陈晓妹[1] 王飞[1] 张翼[1] 刘丽玲[1] 田国斌[1] 曾显营[1] 管雪婷[1] 程成[1] 陈化兰[1] 李雁冰[1] 

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

出　　处：《中国预防兽医学报》2011年第6期439-442,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：农业部行业专项-边境动物疫病防制技术研究(200803026);农业部动物流感防治与监测专项

摘　　要：为制备特异性的H9亚型禽流感病毒(AIV)单因子血清,本研究分别将6株不同亚群的H9亚型AIV的血凝素(HA)基因以鸡偏嗜的密码子进行优化,经全基因合成插入高效真核表达载体pCAGGS中,构建的真核重组质粒转染293T细胞进行瞬时表达,间接免疫荧光试验结果表明,重组质粒中的HA目的基因获得表达。将重组质粒以200μg/只的剂量免疫1月龄SPF鸡,6周后采血分离血清。交叉微量血凝抑制试验结果表明,血凝抑制效价可达8 log2～12 log2,灵敏度高,与其他AIV亚型抗原无交叉反应,型特异性强。To prepare hemagglutinin（HA） antiserum against H9 subtype avian influenza virus（AIV）,the HA gene of 6 different clades of H9 subtype AIV were optimized with chicken＇s codon and chemically synthesized.The codon optimized HA genes were inserted into the eukaryotic expression vector pCAGGS to construct recombinant expression plasmid,which were transfected into 293T cells.The expressed HA proteins were verified by indirect immunofluorescence assay on 48 hours post transfection.One-month-old SPF chickens were immunized with the recombinant plasmid intramuscularly at a dose of 200 μg.The antiserum were collected on weeks 6 post inoculation.The results of cross hemagglutinin inhibition（HI） test demonstrated that the antiserum titer reached from 8 log2 to 12 log2,and no interaction with other AIV subtypes antigen.The antiserum was proved to be high specificity to AIV subtype H9 and could be used as subtype-specific reference antiserum.

关 键 词：禽流感病毒 H9亚型 真核表达重组质粒 单因子血清 

分 类 号：S852.65[农业科学—基础兽医学]