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.

[1]  H. Weingartl,et al.  Corrigendum to "Development of a Rift Valley fever real-time RT-PCR assay that can detect all three genome segments" [J. Virol. Methods 193 (2) (2014) 426-431]. , 2014, Journal of Virological Methods.

[2]  J. Kortekaas One Health approach to Rift Valley fever vaccine development. , 2014, Antiviral research.

[3]  T. Oury,et al.  Broad Spectrum Antiviral Activity of Favipiravir (T-705): Protection from Highly Lethal Inhalational Rift Valley Fever , 2014, PLoS neglected tropical diseases.

[4]  M. Groschup,et al.  A novel indirect ELISA based on glycoprotein Gn for the detection of IgG antibodies against Rift Valley fever virus in small ruminants. , 2013, Research in veterinary science.

[5]  M. Turell,et al.  Potential for Mosquitoes (Diptera: Culicidae) From Florida to Transmit Rift Valley Fever Virus , 2013, Journal of medical entomology.

[6]  T. Mizutani,et al.  Antigen-capture ELISA for the detection of Rift Valley fever virus nucleoprotein using new monoclonal antibodies. , 2012, Journal of virological methods.

[7]  Zengqi Yang,et al.  Development of a RT-PCR for the Detection of European Brown Hare Syndrome Virus , 2011 .

[8]  G. H. Gerdes Rift Valley fever. , 2004, Revue scientifique et technique.

[9]  Z. Memish,et al.  Rift Valley fever: an uninvited zoonosis in the Arabian peninsula. , 2003, International journal of antimicrobial agents.

[10]  Eric Cardinale,et al.  Development of real-time RT-PCR for the detection of low concentrations of Rift Valley fever virus. , 2014, Journal of virological methods.

[11]  M. Turell,et al.  Vector Competence of Selected African Mosquito (Diptera: Culicidae) Species for Rift Valley Fever Virus , 2008, Journal of medical entomology.