表达1型和3型鸭甲型肝炎病毒VP1基因重组新城疫病毒的构建及其免疫原性

发布时间：2018-02-27 13:32

本文关键词： 鸭甲型肝炎病毒(DHAV) DHAV- DHAV- 反向遗传操作重组新城疫病毒　出处：《中国兽医学报》2017年09期 　论文类型：期刊论文

【摘要】：为进一步研发安全有效的鸭甲型肝炎病毒(DHAV)多价疫苗,本研究以新城疫病毒(NDV)LaSota弱毒疫苗株为载体,构建出共表达DHAV-1和-3型VP1基因的重组病毒rLS-1VP1-2A-3VP1,并对其毒力、生物学特性进行鉴定。通过间接免疫荧光试验(IFA)验证DHAV-1和-3型VP1蛋白在细胞内能够有效表达。利用Western blot进一步验证重组病毒产生的中和抗体能和DHAV-1和DHAV-3VP1表达蛋白发生较强的反应。重组病毒rLS-1VP1-2A-3VP1保持了原重组疫苗株rLS-RFP对鸡胚的高滴度生长适应和低致病的特性。将重组病毒rLS-1VP1-2A-3VP1免疫种鸭1次,通过中和试验检测该重组病毒能够在鸭体内产生针对DHAV-1和DHAV-3的中和抗体,平均效价分别为1∶18.17和1∶16.60。本研究结果表明重组病毒rLS-1VP1-2A-3VP1具有作为鸭肝炎重组病毒活载体疫苗用于种鸭的潜力。
[Abstract]:In order to further develop a safe and effective duck hepatitis A virus (DHAV) multivalent vaccine, the recombinant virus rLS-1VP1-2A-3VP1 was constructed and its virulence was studied by using NDV (NDV) LaSota attenuated vaccine strain of Newcastle disease virus (NDV) as vector, and the recombinant virus rLS-1VP1-2A-3VP1 was constructed. By indirect immunofluorescence assay (IFA), DHAV-1 and type -3 VP1 proteins were effectively expressed in cells. Western blot was used to further verify the neutralizing antibody produced by the recombinant virus and DHAV-1 and DHAV-3VP1 expression proteins. The recombinant virus rLS-1VP1-2A-3VP1 maintained the characteristics of high titer growth adaptation and low pathogenicity of the original recombinant vaccine strain rLS-RFP to chicken embryo. The recombinant virus rLS-1VP1-2A-3VP1 was immunized with duck once. Neutralization tests showed that the recombinant virus produced neutralizing antibodies against DHAV-1 and DHAV-3 in ducks. The average titers were 1: 18.17 and 1: 16.60 respectively. The results showed that the recombinant virus rLS-1VP1-2A-3VP1 had the potential to be used as live vector vaccine for duck hepatitis virus.
【作者单位】：山东农业大学动物科技学院;山东省农业科学院家禽研究所;
【基金】：山东省现代农业产业技术体系资助项目(SDAIT-13-011-01) 山东省农业科学院青年科研基金资助项目(2014QNZ06)
【分类号】：S852.65

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