Design and Evaluation of the Primers for Rift Valley Fever (RVF) Virus RT-PCR Detection

Rift Valley Fever (RVF) is a notifiable multiple species diseases in the OIE list, and causes human and agricultural losses in endemic regions. To develop the rapid method for detecting of RVF, 2 specific primers for reverse transcriptase polymerase chain reaction (RT-PCR) and 7 overlapping oligo primers were designed according to the nucleotide sequence information of RVFV published in GenBank, and a DNA fragment about 318 bp of the segment S was synthesized in vitro by overlap extension PCR to construct the recombinant plasmid pMD19-T-RVFVS. Then, the 2 specific primers were evaluated via a serial of tests, including reaction temperature optimization test, sensitivity and specificity tests. The results showed that the 2 designed primers are suitable for RVFV RT-PCR detection which is a rapid method with good specificity and sensitivity, the detection limit was approximately 85 copies of the cloned viral genomic fragments (pMD19-T-RVFVS) as well as resulted in no cross-reaction for peste des petits ruminants virus (PPRV), Epidemic encephalitis B virus, E.coli , Salmonella and Pasteurella multocida etc common pathogens isolated from ruminants detection.