Bluetongue (BT) is non-contagious, vector-borne viral disease of domestic and wild ruminants, transmitted by midges (Culicoides spp.) and is caused by Bluetongue virus (BTV). BTV is the type species of the Orbivirus genus within the Reoviridae family and possesses a genome consisting of 10 double-stranded RNA segments encoding 7 structural and 4 nonstructural proteins. Viral Protein 7 (VP7) is the major sera group-specific protein and is a good antigen candidate for immunoenzymatic assays for the BT diagnosis. In our work, BTV-2 recombinant VP7 (BTV-2 recVP7), expressed in Spodoptera frugiperda (Sf9) cells using a baculovirus system, was produced and purified by affinity chromatography from the supernatant of infected cell culture. The use of the supernatant allowed us to obtain a high quantity of recombinant protein with high purity level by an easy one-step procedure, rather than the multistep purification from the pellet. RecVP7-BTV2 was detected using a MAb anti-BTV in Western blot and it was used to develop an immunoenzymatic assay.

蓝舌病 (BT) 是家养和野生反刍动物的非传染性病媒传播病毒性疾病，由蠓 ( Culicoides spp.) 传播，由蓝舌病毒 (BTV) 引起。BTV 是呼肠孤病毒科中环状病毒属的模式种，其基因组由 10 个双链 RNA 片段组成，编码 7 个结构蛋白和 4 个非结构蛋白。病毒蛋白 7 (VP7) 是主要的血清组特异性蛋白，是用于 BT 诊断的免疫酶测定的良好抗原候选者。在我们的工作中，在草地贪夜蛾中表达的 BTV-2 重组 VP7 (BTV-2 recVP7)(Sf9) 细胞使用杆状病毒系统，通过亲和层析从感染细胞培养物的上清液中产生和纯化。上清液的使用使我们能够通过简单的一步程序获得大量高纯度的重组蛋白，而不是从沉淀中进行多步纯化。RecVP7-BTV2 在蛋白质印迹中使用 MAb 抗 BTV 进行检测，并用于开发免疫酶测定。