【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19.2μg/mL,最佳包被条件为37℃1h后4℃过夜;血清的最佳稀释度为1∶800,37℃孵育60min;酶标二抗稀释度为1∶7 000,37℃孵育60min;底物显色为37℃避光作用10min。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。更多还原

【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 19.2μg/mL and the optimal condition for coating was incubated at 37℃for 1hand then 4℃overnight.The optimal dilutions of serum and enzyme labeled antibody were 1∶800and 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.更多还原