Bluetongue (BT) is an arbovirus-borne disease of ruminants caused by bluetongue virus (BTV) that has the potential to have a serious economic impact. Currently available commercial vaccines include attenuated vaccines and inactivated vaccines, both of which have achieved great success in the prevention and control of BTV. However, these vaccines cannot distinguish between infected animals and immunized animals. To control outbreaks of BTV, the development of labeled vaccines is urgently needed. In this study, we used the plasmid-based reverse genetics system (RGS) of BTV to rescue four recombinant viruses in which HA (influenza hemagglutinin) tags were inserted at different sites of VP2. In vitro, the recombinant tagged viruses exhibited morphologies, plaque, and growth kinetics similar to the parental BTV-16, and expressed both VP2 and HA tag. Subsequently, the selected recombinant tagged viruses were prepared as inactivated vaccines to immunize IFNAR(−/−) mice and sheep, and serological detection results of anti-HA antibody provided discriminative detection. In summary, we used plasmid-based RGS to rescue BTV recombinant viruses with HA tags inserted into VP2, and detected several sites on VP2 that can accommodate HA tags. Some of the recombinant tagged viruses have potential to be developed into distinctive inactivated vaccines.

蓝舌病（BT）是虫媒病毒传播的反刍动物疾病，由蓝舌病病毒（BTV）引起，具有潜在的严重经济影响。当前可用的商业疫苗包括减毒疫苗和灭活疫苗，两者在预防和控制BTV方面均取得了巨大成功。但是，这些疫苗不能区分感染的动物和免疫的动物。为了控制BTV的爆发，迫切需要开发标记疫苗。在这项研究中，我们使用BTV的基于质粒的逆向遗传系统（RGS）抢救了四种重组病毒，其中HA（流感血凝素）标签插入了VP2的不同位点。体外，重组标记的病毒表现出类似于亲本BTV-16的形态，噬菌斑和生长动力学，并表达VP2和HA标签。随后，将选择的重组标签病毒制备为灭活疫苗，以免疫IFNAR（-/-）小鼠和绵羊，抗HA抗体的血清学检测结果提供了判别性检测。总而言之，我们使用了基于质粒的RGS来拯救带有在VP2中插入HA标签的BTV重组病毒，并在VP2上检测到了几个可以容纳HA标签的位点。一些重组标记的病毒有潜力被开发成独特的灭活疫苗。