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H9亚型禽流感病毒血凝素特异性单因子血清制备
引用本文:陈晓妹,王飞,张翼,刘丽玲,田国斌,曾显营,管雪婷,程成,陈化兰,李雁冰.H9亚型禽流感病毒血凝素特异性单因子血清制备[J].中国预防兽医学报,2011,33(6).
作者姓名:陈晓妹  王飞  张翼  刘丽玲  田国斌  曾显营  管雪婷  程成  陈化兰  李雁冰
作者单位: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
基金项目:农业部行业专项,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 long2~12 log2,灵敏度高,与其他AIV亚型抗原无交叉反应,型特异性强.

关 键 词:禽流感病毒  H9亚型  真核表达重组质粒  单因子血清

Preparation of hemagglutinin antiserum against H9 subtype avian influenza virus
CHEN Xiao-mei,WANG Fei,ZHANG Yi,LIU Li-ling,TIAN Guo-bin,ZENG Xian-ying,GUAN Xue-ting,CHENG Cheng,CHEN Hua-lan,LI Yan-bing.Preparation of hemagglutinin antiserum against H9 subtype avian influenza virus[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(6).
Authors:CHEN Xiao-mei  WANG Fei  ZHANG Yi  LIU Li-ling  TIAN Guo-bin  ZENG Xian-ying  GUAN Xue-ting  CHENG Cheng  CHEN Hua-lan  LI Yan-bing
Institution:CHEN Xiao-mei,WANG Fei,ZHANG Yi,LIU Li-ling,TIAN Guo-bin,ZENG Xian-ying,GUAN Xue-ting,CHENG Cheng,CHEN Hua-lan*,LI Yan-bing*(Animal Influenza Laboratory of the Ministry Agricultural,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China)
Abstract: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 im...
Keywords:avian influenza virus  H9 subtype  eukaryotic expression recombinant plasmid  hemagglutinin subtype-specific reference antiserum  
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