A Species-Independent Lateral Flow Test to Detect Rift Valley Fever Virus Antibodies Using a Double Antigen Approach

Rift Valley fever virus (RVFV) is a re-emerging, vector-borne pathogen endemic to Africa and the Arabian Peninsula, posing an increasing threat to human and animal health. Outbreaks have severe economic and social impacts on farmers, communities, and governments. Current diagnostic methods rely on PCR and ELISA; however, rapid pen-side tests would enable faster, cost-effective monitoring and outbreak control. Here, a species- and immunoglobulin class-independent capillary flow immunodiagnostic assay (lateral flow test; LFT) for detecting RVFV-specific antibodies is described. The assay uses a double-antigen approach, coupling the RVFV nucleocapsid protein, a major viral antigen, both to carbon nanoparticles and to a nitrocellulose membrane. The method was qualified with immune sera from sheep, calves, goats, and humans and benchmarked against a newly developed double-antigen ELISA and a commercial competition ELISA. Both the LFT and double-antigen ELISA demonstrated high specificity and sensitivity. This advancement brings RVFV-specific pen-side testing significantly closer to practical implementation.