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禽腺病毒12个血清型代表毒株的分子生物学鉴定研究
侯力丹,宋佳诚,刘丹,毛娅卿,杨亚希,黄小洁,吴应凯,王嘉,李俊平
0
(中国兽医药品监察所)
摘要:
禽腺病毒(FAdV)分为5个种,12个血清型,多个种和血清型的特征为FAdV的鉴定、检验及诊断方法建立和相关疫苗评价等带来很大挑战,背景清晰、纯净特异的12个血清型参考毒株可为FAdV系统鉴定和相关生物制品研发和评价提供重要的保障。本研究对中国兽医药品监察所保藏的FAdV 12个血清型代表毒株进行系统分型鉴定,明确其遗传背景、血清型信息及纯净性,以建立和完善系统完整的FAdV标准毒株库。为此,本研究针对FAdV不同种的Fiber和Hexon序列设计引物,经PCR扩增和遗传进化分析,完成对毒株的分型。再结合同一种中不同血清型病毒Hexon序列上酶切位点的特征,用酶切法验证了毒株不存在同一种中不同血清型之间的交叉污染。综上分析,结合遗传进化分析和酶切法可以实现对FAdV 12个血清型毒株的特异性和纯净性鉴定,本研究建立了完整的FAdV的代表毒株毒种库,为该病检测方法的建立和疫苗研发工作奠定了物质基础。
关键词:  禽腺病毒  代表毒株  分子生物学
DOI:
投稿时间:2021-03-03修订日期:2021-06-10
基金项目:国家重点研发计划“畜禽重大疫病防控与高效安全养殖综合技术研发”重点专项(2016YFD0500800, 2016YFD0500809)
Molecular biological identification of 12 strains of Fowl Adenovirus
(China Institute of Veterinary Drug Control,Beijing)
Abstract:
Fowl adenovirus (FAdV) is divided into 5 types and 12 serotypes. The characteristics of multiple types and serotypes pose great challenges for the identification, testing and diagnosis of FAdV and related vaccine evaluation. The pure and specific serotype reference strains are of vital importance for FAdV identification and development of related biological product. In this study, we attempted to systematically identify the 12 representative serotypes of FAdV preserved by the Institute of Veterinary Drug Control, and to clarify their genetic background, serotype information and purity, in order to establish and improve the complete FAdV standard virus strain library. This study designed primers for the Fiber and Hexon sequences of different species of FAdV, and analyzed the strains by PCR amplification and genetic evolution analysis. In consideration of the characteristics of the restriction sites on the Hexon sequence of different viruses of the same serotype, the enzyme digestion method was used to verify that there was no cross-contamination of other strains in the same serotype. Based on above analysis, combined with genetic evolution analysis and enzyme digestion method, the specificity and purity of the 12 serotype strains of FAdV can be identified. In this study, a complete representative virus strain library of FAdV was established, laying the material foundation of the disease detection method establishment and vaccine research.
Key words:  Avian adenovirus  Representative strain  Molecular biology

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