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4型禽腺病毒抗体间接hexon-ELISA检测方法的建立
朱庆贺,张鹏宇,崔红玉,王观悦,王爽,陈曦,史同瑞
0
(黑龙江省兽医科学研究所)
摘要:
以4型禽腺病毒(FAdV-4)的DNA为模板,扩增其六邻体(hexon)部分基因并进行重组表达,以重组hexon蛋白为包被抗原,优化ELISA 检测条件,建立了FAdV-4的ELISA 抗体检测方法。该方法对FAdV-4阳性血清检测为阳性,对于其他鸡常见病毒,如禽流感病毒5、7、9型,新城疫病毒以及鸡传染性支气管炎病毒阳性血清检测均为阴性;与PCR方法的阳性符合率为83.3%。试验表明,建立的以重组hexon蛋白为包被抗原检测FAdV-4血清抗体的ELISA方法可以用于检测FAdV-4的感染及相关流行病学调查。
关键词:  4型禽腺病毒  hexon蛋白  原核表达  间接ELISA
DOI:
投稿时间:2017-12-27修订日期:2018-07-04
基金项目:黑龙江省应用技术研究与开发计划(sczzn2017-2)
Establishment of an Indirect hexon-ELISA Method for the Detection of Antibodies against Fowl Avianadenovirus Serotype 4
(Heilongjiang Institute of Veterinary Science,Heilongjiang Qiqihar)
Abstract:
FAdV-4 DNA as templates was used to amplify the hexon gene, and the recombinant hexon protein was expressed. With the recombinant hexon protein as the detection antigen, an indirect ELISA was established by optimizing the ELISA detection conditions. FAdV-4 antibodies reacted with the hexon protein significantly, no cross-reactivity occurred between the hexon protein and antibodies against other chicken viruses, such as avian influenza virus (AIV-5,AIV-7,AIV-9), newcastle disease virus (NDV), and infection bronchitis virus (IBV). The positive coincidence rate with PCR method was 83.3%. It showed that the developed ELISA could be used for the diagnosis of FAdV-4 infection and investigation of related epidemiological.
Key words:  FADV-4  hexon protein  prokaryotic  indirect ELISA

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