Abstract: The Large (L) RNA segment of Rift Valley Fever (RVF) virus (RVFV) was evaluated for detection of the virus in serum samples from acute hemorrhagic fever cases using RT-PCR. The RT-PCR produced a 342 base pair (bp) from RVF RNAs samples extracted from sera from infected humans. However, the RT-PCR assay did not amplify the specific 342 bp product from RNAs extracted from closely related hemorrhagic fever viruses including Crimean Congo Hemorrhagic Fever Virus (CCHFV), Dengue virus, total RNA extracted from non infected vero cells. The described RT-PCR provides rapid, sensitive and specific method for detection of RVFV RNA directly in human sera sampled during acute phase of the disease.