Introduction

Viral hemorrhagic fevers (VHFs) encompass a group of infectious diseases that interfere with the body’s ability to clot blood, often leading to serious complications including death. The clinical presentation of VHF’s are non-specific, making it challenging to distinguish by symptoms alone, especially from other infections like Malaria that may be co-circulating. There is a need to correctly detect and differentiate these infections, to ensure the correct treatments and control measures are implemented. Multiplex nucleic acid tests enable the screening of multiple targets at once, with techniques like recombinase polymerase amplification (RPA) allowing for rapid detection without the need for complex instrumentation. However, RPA assays are often constricted by the degree of multiplexing that can be carried out concurrently per reaction, while the complex RNA secondary structure has been known to reduce the sensitivity of a one-step RT-RPA reaction.