| 鸡传染性支气管炎病毒抗体间接ELISA检测方法的建立 |
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| 引用本文： | 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.鸡传染性支气管炎病毒抗体间接ELISA检测方法的建立[J].中国畜牧兽医,2015,42(6):1383-1388. |
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| 作者姓名： | 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 |
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| 作者单位： | 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 |
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| 基金项目： | 山西省科技攻关项目(20120311023-5) |
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| 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.
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| 关 键 词： | 鸡传染性支气管炎病毒(IBV) 间接ELISA 抗体检测 |
| 收稿时间： | 2014-11-10 |
Establishment of Indirect ELISA Method for Detecting Antibodies against Avain Infectious Bronchitis Virus |
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| 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.Establishment of Indirect ELISA Method for Detecting Antibodies against Avain Infectious Bronchitis Virus[J].China Animal Husbandry & Veterinary Medicine,2015,42(6):1383-1388. |
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| Authors: | 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 |
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| Institution: | Veterinary Microbiology and Immunology Laboratory, College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China |
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| 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. |
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| Keywords: | avian infectious bronchitis virus (IBV) indirect ELISA antibody detection |
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