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I群禽腺病毒4型IgG抗体间接ELISA检测方法的建立
张启龙,孙丹,韦海涛,宋彦军,周德刚,冯小宇,王林
0
(北京市动物疫病预防控制中心)
摘要:
为建立I群禽腺病毒4型血清学检测方法,本研究利用纯化的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1 μg/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60min;酶标抗体的最佳作用条件为稀释度1:8000,37℃孵育60min;最佳显色时间为10min。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。
关键词:  I群禽腺病毒4型  Hexon蛋白  间接ELISA  检测
DOI:
投稿时间:2019-05-06修订日期:2019-05-28
基金项目:
Establishment of an Indirect ELISA for theDetection of IgG Antibody of Group I Fowl Adenovirus Serotype4
(Beijing Center for Animal Disease Control and Prevention)
Abstract:
In order to establish a serological method for detection of group I fowl adenovirus serotype4,under screening various reaction conditions, an indirect ELISA was established for detection of group I fowl adenovirus serotype4 IgG antibody by used 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 conditons 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.
Key words:  Group I Fowl adenovirus serotype4  Hexon protein  Indirect ELISA  Detection

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