Molecular Diagnosis of Phlebovirus riftense Infection Using an L-Segment-Based Real-Time RT-PCR Method

Rift Valley fever (RVF) is one of the main vector-borne zoonotic diseases that affects a wide range of ruminants and humans in Africa and the Arabian Peninsula.Several techniques involving cell culture and molecular biology methods have been developed to diagnose RVF infection. Success partly relies on sending samples to a national or reference laboratory in good conditions and having the capacity to perform the appropriate diagnostic test in the matrix of interest during the period of viremia where high loads of viral particles are present. Conventional and duplex/multiplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) assays are currently the most rapid and sensitive molecular tests for the detection and quantification of Rift Valley fever virus (RVFV) during outbreaks. The one-step real-time duplex RT-PCR technique that is detailed in this chapter is based on the L segment of RVFV. Results must be carefully validated and interpreted, taking into account the results of both positive and negative controls.