Expression of Foot-and-Mouth Disease Virus Non-Structural Protein, 3D in Insect Cells and its Application in Detection of Anti-FMDV Antibodies.

Indian Journal of Virology Pub Date : 2012-12-01 Epub Date: 2012-09-06 DOI:10.1007/s13337-012-0098-8
Rakesh Kumar, M Hosamani, B P Sreenivasa, Anil Kotyal, R Venkataramanan
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引用次数: 12

Abstract

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.

口蹄疫病毒非结构蛋白3D在昆虫细胞中的表达及其在检测口蹄疫病毒抗体中的应用
基于非结构蛋白(NSPs)的诊断对于口蹄疫(FMD)的大规模血清监测和监测病毒活性是有用的,作为口蹄疫流行国家(如印度)旨在通过疫苗接种控制疾病的疫苗接种运动的后续行动。这些诊断方法在进出口偶蹄动物的血清学中也很方便。本研究利用杆状病毒表达系统对口蹄疫病毒(FMDV)非结构蛋白RNA聚合酶(3D基因)进行了表达。用SDS-PAGE分析蛋白表达,并用western blot检测其与感染口蹄疫病毒的牛血清的免疫反应性。用1072份牛血清样品纯化重组3D蛋白并进行间接ELISA检测。当对已知口蹄疫状态的牛血清进行检测时,发现该检测方法的诊断敏感性和特异性分别为92%和100%。本文开发的基于3D的ELISA可用于动物筛选,作为其他基于NSP的诊断方法的辅助手段,可用于口蹄疫常规血清监测。
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来源期刊
Indian Journal of Virology
Indian Journal of Virology 医学-病毒学
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6-12 weeks
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