畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (5): 679-684.doi:

• 预防兽医 • 上一篇    下一篇

马流感病毒双抗体夹心ELISA检测方法的建立

姬媛媛,郭巍,王晓钧,王征,卢刚,赵立平,李红梅,相文华*   

  1. 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
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-05-26 发布日期:2011-05-26
  • 通讯作者: 相文华

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

JI Yuanyuan, GUO Wei, WANG Xiaojun, WANG Zheng, LU Gang,
ZHAO Liping, LI Hongmei, XIANG Wenhua﹡   

  • Received:1900-01-01 Revised:1900-01-01 Online:2011-05-26 Published:2011-05-26

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。

Abstract: 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.