【摘要】 利用纯化的Ⅰ群禽腺病毒4型重组Hexon蛋白,通过各反应条件筛选,建立了Ⅰ群禽腺病毒4型IgG抗体间接ELISA检测方法。结果显示,抗原最佳包被浓度为1μg/mL;最佳封闭液为1%牛血清白蛋白;待检血清最佳作用浓度是400倍稀释,37℃孵育60 min;酶标抗体的最佳作用条件为稀释度1∶8000,37℃孵育60 min;最佳显色时间为10 min。用所建方法对160只临床鸡只进行检测,同时与拭子PCR检测对比,一致性较好,表明该方法可有效用于临床检测。更多还原

【Abstract】 With various reaction conditions being screened,an indirect ELISA was established for detection of group Ⅰ Fowl adenovirus serotype 4 IgG antibody by using the purified recombination Hexon protein. The results showed that the optimal concentration of the coating antigen was 1 μg/mL; the best blocking solution was 1% bovine serum albumin; the optimal concentration of the serum to be tested was 400-fold dilution,and the optimal incubation time of the serum was 60 minutes at 37 ℃; the optimal conditions of the enzyme-labeled antibody were 1 ∶8000 dilution,and incubated for 60 minutes at 37 ℃; the optimal chromogenic time was 10 minutes. The established method was used to detect 160 clinical chickens and compared with the swab PCR test,the results were consistent,indicating that the proposed method can be used for clinical detection effectively.更多还原