马流感病毒双抗体夹心ELISA检测方法的建立  

Development of Double Antibody Sandwich ELISA for Detection of Equine Influenza Virus

作　　者：姬媛媛[1] 郭巍[1] 王晓钧[1] 王征[2] 卢刚[3] 赵立平[1] 李红梅[1] 相文华[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物病研究室,哈尔滨150001 [2]东北农业大学生命科学学院,哈尔滨150030 [3]黑龙江八一农垦大学食品学院,大庆163319

出　　处：《畜牧兽医学报》2011年第5期679-684,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：黑龙江省自然科学基金重点项目(ZD200812)

摘　　要：为建立一种快速、有效的检测马流感病毒(Equine influenza virus,EIV)的方法,以EIV中国分离株A/马/新疆/07(H3N8)制备的多克隆抗体为捕获抗体,原核表达的核蛋白(NP)制备的单克隆抗体为检测抗体,在国内首次建立了检测EIV的双抗体夹心ELISA方法。用该检测方法分别检测EIV、马动脉炎病毒、马疱疹病毒1型、马疱疹病毒4型和马乙型脑炎病毒阳性样品。结果表明,该ELISA方法具有良好的特异性;与常规检测EIV的血凝试验相比,其敏感性是后者的2.5～10倍;同时与H7N7亚型EIV有交叉反应。攻毒试验结果表明该方法可有效检测鼻腔分泌物中的EIV。该方法的建立为EIV的检测及早期防控提供了有效工具。To develop a rapid and effective method for Equine influenza virus（EIV） detection,polyclonal antibodies against EIV A/equine/Xinjiang/07 strain and monoclonal antibody against NP of EIV were generated respectively.Then a double antibody sandwich ELISA（DAS-ELISA） was developed.The specificity of the optimized DAS-ELISA was evaluated using EIV,Equine arteritis virus,Equine herpes virus-1,Equine herpes virus-4 and Japanese encephalitis virus,resulting in only EIV specimens yielding a strong signal.Compared with hemagglutination test,its sensitivity was as two point five to ten times as the later.And it had cross-reactivity with H7N7 subtype.Meanwhile it is suitable for detection of virus from the nasal swabs of experimentally infected equines.The results revealed that the ELISA possessed good specificity and higher sensitivity,indicating a suitable method for rapid detection of EIV.

关 键 词：马流感病毒 多克隆抗体 单克隆抗体 双抗体夹心ELISA 

分 类 号：S852.659[农业科学—基础兽医学] S852.652[农业科学—兽医学]