Development of a Quantitative RT-PCR Assay to Differentiate Rift Valley Fever Virus Smithburn Vaccine Strain from Clone 13 Vaccine Strain.

Abstract

A new quantitative RT-PCR assay was developed to differentiate Rift Valley fever (RVF) Smithburn vaccine strain from Clone 13 vaccine strain. The new qRT-PCR assay targeting the S segment (NSs and N gene) was tested on synthesized standard RNA and MP-12 strain viruses. The detection limit of the new qRT-PCR assay is 1 copy/μL of NSs and N, and is able to differentiate the Smithburn vaccine strain of RVF from the Clone 13 vaccine strain. No cross-reactivity with other vector-borne viruses was observed, a factor that is especially important in the Republic of Korea (ROK). To examine the performance of the qRT-PCR, intra- and inter-assay variability data were analyzed and showed high reproducibility. These results indicate that the new qRT-PCR can be used as a safe and cost-effective test. Furthermore, this result suggests the possibility of differentiation between infected and vaccinated animals diagnostic test in RVF-free countries including ROK.