Protein-based tools for the detection and characterisation of Oropouche virus infection

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.