Merchant Monique K, de Paula Souza Juliano, Abdelkarim Sahar, Chakraborty Shrestha, Nasser Neto Tufi A, Gutierrez Manchay Kristel I, de Melo Jorge Daniel M, do Nascimento Valdinete Alves, Sun Yuxian, Caroe Eve R, Eyssen Lauren E A, Rudd Jared S, Ribeiro Piauilino Isa C, Damasceno Pinto Sérgio, Pereira da Silva Matheus H, Rocha do Nascimento Felipe, Naveca Felipe Gomes, Proenca-Modena José L, da Silva Luis L P, Pinto de Figueiredo Regina M, Owens Raymond J, Arruda Eurico, Graham Stephen C
Department of Pathology, University of Cambridge, Cambridge, CB2 1QP, UK.
Department of Cell Biology and Center for Virus Research, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, 14049-900, Brazil.
EMBO Mol Med. 2025 Aug 11. doi: 10.1038/s44321-025-00291-7.
Oropouche fever is a neglected tropical disease caused by the orthobunyavirus Oropouche virus (OROV). A recent OROV epidemic caused by a novel reassortant has seen infections across an expanded geographical range, with deaths of healthy adults plus vertical transmission leading to pregnancy loss. OROV research and epidemiology is hampered by a paucity of available tools for serology and molecular virology. We have purified recombinant OROV nucleoprotein and the spike region of the viral surface glycoprotein Gc. These antigens detect seroconversion following experimental infection of animals in indirect ELISA, confirming their antigenic authenticity. They stimulate the production of high neutralising antibody titres in animals, highlighting their promise as immunogens for vaccination. We developed a nanobody-based sandwich ELISA that can detect OROV antigens in human clinical serum samples with high efficiency, and we show that nanobodies directed against OROV Gc spike can potently neutralise infection by both historical OROV strains and the newly emerged reassortant. Our protein-based reagents will accelerate OROV research and highlight the utility of protein-based tools for future OROV vaccines and point-of-care diagnostic devices.
奥罗普切热是一种由正布尼亚病毒奥罗普切病毒（OROV）引起的被忽视的热带疾病。最近由一种新型重配病毒引起的OROV疫情在更广泛的地理范围内出现了感染病例，包括健康成年人死亡以及垂直传播导致流产。OROV的研究和流行病学受到血清学和分子病毒学可用工具匮乏的阻碍。我们已经纯化了重组OROV核蛋白以及病毒表面糖蛋白Gc的刺突区域。这些抗原在间接ELISA中可检测到动物实验感染后的血清转化，证实了它们的抗原真实性。它们能刺激动物产生高中和抗体滴度，凸显了它们作为疫苗免疫原的潜力。我们开发了一种基于纳米抗体的夹心ELISA，可高效检测人类临床血清样本中的OROV抗原，并且我们表明针对OROV Gc刺突的纳米抗体能够有效中和历史OROV毒株和新出现的重配病毒的感染。我们基于蛋白质的试剂将加速OROV研究，并突出基于蛋白质的工具在未来OROV疫苗和即时诊断设备中的实用性。
