Development of RT-PCR for Detection of Rift Valley Fever Virus Based on the Medium (M) Gene Segment of the Virus

Abstract

In the present study, reverse transcriptase polymerase chain reaction RT-PCR protocol was carried out to detect Rift Valley fever virus (RVFV) RNA in Vero cell culture and serum samples. The RVFV vaccine strain smith burn and sera from infected humans were used in this study. The viruses were propagated in Vero cell culture. RNAs were extracted from cell culture and directly from serum samples and then, they were detected by the described RT-PCR assay, using primers derived from medium (M) RNA segment of the virus. The specific 342-bp PCR products were amplified from RVFV infected cell culture and serum samples, and they were visualized on ethidium bromide-stained agarose gel. The ampilification product was not detected when the RT-PCR assay was applied to RNA from Crimean Congo hemorrhagic fever virus (CCHFV), Dengue and total RNA extracted from non infected Vero cells. The 342-bp specific PCR product was detected from 1.0 pg of RNA target from smith burn vaccine strain and sera from infected humans. The result of this study showed that the described RT-PCR assay, using the well characterized primers, could be applied for detection of RVFV from cell culture and clinical samples.