首页> 美国卫生研究院文献>Indian Journal of Virology >Expression of Foot-and-Mouth Disease Virus Non-Structural Protein 3D in Insect Cells and its Application in Detection of Anti-FMDV Antibodies
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Expression of Foot-and-Mouth Disease Virus Non-Structural Protein 3D in Insect Cells and its Application in Detection of Anti-FMDV Antibodies

机译:口蹄疫病毒非结构蛋白3D在昆虫细胞中的表达及其在抗FMDV抗体检测中的应用

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摘要

Non-structural proteins (NSPs) based diagnostics are useful for large-scale sero-surveillance of foot-and-mouth disease (FMD) and to monitor viral activity as a follow up to the vaccination campaign in FMD endemic countries like India which aim at disease control through vaccination. These diagnostics are also handy in the serology of import/export of cloven-footed animals. In the present study, non-structural protein RNA polymerase (3D gene) of FMD virus (FMDV) was expressed using baculovirus expression system. Protein expression was analyzed by SDS-PAGE and confirmed by its immuno-reactivity with serum from a FMDV infected bovine, in the western blot. Recombinant 3D protein was purified and evaluated in the indirect ELISA with 1072 cattle serum samples. Diagnostic sensitivity and specificity of the assay were found to be 92 and 100 %, respectively, when tested with cattle sera of known FMD status. The 3D based ELISA developed here is useful for screening the animals as an adjunct to other NSP based diagnostics available for routine serosurveillance of FMD.
机译:基于非结构蛋白(NSP)的诊断可用于口蹄疫(FMD)的大规模血清监测以及监测病毒活动,作为针对印度等口蹄疫流行国家的疫苗接种运动的后续行动通过疫苗控制疾病。这些诊断在丁香动物进出口的血清学检查中也很方便。在本研究中,使用杆状病毒表达系统表达了口蹄疫病毒(FMDV)的非结构蛋白RNA聚合酶(3D基因)。通过SDS-PAGE分析蛋白质表达,并通过蛋白质印迹法对其与来自FMDV感染的牛血清的免疫反应性进行确认。纯化重组3D蛋白,并在间接ELISA中对1072头牛血清样品进行评估。用已知FMD状态的牛血清测试时,该方法的诊断敏感性和特异性分别为92%和100%。此处开发的基于3D的ELISA可用于筛选动物,作为可用于FMD常规血清监测的其他基于NSP的诊断的辅助工具。

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