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.