《中国畜牧兽医》---唯一指定的官方网站 ›› 2015, Vol. 42 ›› Issue (6): 1383-1388.doi: 10.16431/j.cnki.1671-7236.2015.06.009

• 生物技术 • 上一篇    下一篇

鸡传染性支气管炎病毒抗体间接ELISA检测方法的建立

郑卫峰, 闫芳, 王盼, 赵宇军, 高文伟, 马海利, 李绪英, 宁官保, 田文霞, 梁慧霞, 刘林   

  1. 山西农业大学动物科技学院, 兽医微生物与免疫学实验室, 太谷 030801
  • 收稿日期:2014-11-10 出版日期:2015-06-20 发布日期:2015-07-23
  • 通讯作者: 闫芳 E-mail:yanfang6615@163.com
  • 作者简介:郑卫峰(1991-),男,山西晋城人,硕士生,研究方向:动物传染病的诊断与防治,E-mail:zwf412261484@126.com
  • 基金资助:

    山西省科技攻关项目(20120311023-5)

Establishment of Indirect ELISA Method for Detecting Antibodies against Avain Infectious Bronchitis Virus

ZHENG Wei-feng, YAN Fang, WANG Pan, ZHAO Yu-jun, GAO Wen-wei, MA Hai-li, LI Xu-ying, NING Guan-bao, TIAN Wen-xia, LIANG Hui-xia, LIU Lin   

  1. Veterinary Microbiology and Immunology Laboratory, College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China
  • Received:2014-11-10 Online:2015-06-20 Published:2015-07-23

摘要: 为建立快速检测鸡传染性支气管炎病毒(IBV)的血清学方法,本试验以IBV为检测抗原,建立了一种检测IBV抗体的间接ELISA方法,并对各种检测条件进行了优化.研究结果显示,抗原的最佳包被浓度为19.2 μg/mL,最佳包被条件为37 ℃ 1 h后4 ℃过夜;血清的最佳稀释度为1:800,37 ℃孵育60 min;酶标二抗稀释度为1:7 000,37 ℃孵育60 min;底物显色为37 ℃避光作用10 min.经特异性、重复性、敏感性试验证明,该方法与鸡常见病原的阳性血清均无交叉反应,重复性较好及血清稀释至1:12 800时仍可检测到IBV抗体.结果表明,本试验所建立的间接ELISA方法具有良好的特异性、重复性和敏感性.

关键词: 鸡传染性支气管炎病毒(IBV); 间接ELISA; 抗体检测

Abstract: The purpose of this research was to establish a rapid detection serological method against avain infectious bronchitis virus (IBV).In this study,an indirect ELISA method was established using IBV as the detected antigen and a variety of testing conditions were optimized.The results showed that the optimal antigen coating concentration was 19.2 μg/mL and the optimal condition for coating was incubated at 37 ℃ for 1 h and then 4 ℃ overnight.The optimal dilutions of serum and enzyme labeled antibody were 1:800 and 1:7 000 incubated at 37 ℃ for 60 min,respectively.The optimal condition of chromogenic substrate was incubated at 37 ℃ for 10 min in the dark.The specificity,repeatability and sensitivity tests proved that the indirect ELISA did not cross-react with positive antiviral sera of other chicken diseases,had good repeatability and could detect IBV antibody when serum was diluted 1:12 800.We concluded that the established indirect ELISA was specific,repeatable and sensitive.

Key words: avian infectious bronchitis virus (IBV); indirect ELISA; antibody detection

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