A single-tube RT-PCR amplification for detection of Rift Valley Fever Virus

Abstract

A reverse Transcriptase (RT) Polymerase Chain Reaction (RT-PCR) was developed and evaluated for detection of Rift valley fever virus in cell culture and serum samples from acute hemorrhagic fever cases. A pair of primers selected from the Medium (M) RNA gene segment of RVFV was used for RT-PCR amplification. The RT-PCR produced a 342-base pair (bp) from RVF RNAs extracted from smith burn vaccine strain propagated in vero cell culture and sera from infected humans and animals. 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 in cell culture and directly in serum samples. The RT-PCR assay should be recommended for inclusion during epidemiological surveys and outbreaks of the disease among humans and susceptible animal populations. © Medwell Journals, 2010.