Per Signature Limit of Detection for Rift Valley Fever Strains Tested: All tests were conducted on total nucleic acid extracts from clarified viral cultures containing the listed strains.

QRT-PCR tests were performed in triplicate on the ABI7500FAST platform as described in materials and methods. An 8-log dilution series of templates ranging from 200 pfu/ul to 2×10−4 pfu/ul (or 20 pg/ul to 2×10−6 pg/ul for MP-12) was made for each template. Five microliters of each dilution was spiked into qRT-PCR plates in triplicate.*A plaque-forming unit (PFU) is a measure of the number of particles capable of forming plaques per unit volume, such as virus particles. It is a functional measurement rather than a measurement of the absolute quantity of particles: viral particles that are defective or which fail to infect their target cell will not produce a plaque and thus will not be counted.**For RVF MP-12, no titer information is provided as multiple attempts to titer this species failed due to lack of consistent CPE. Thus, sensitivity of this assay is reported in pg of total nucleic acid extract from viral culture. No other RNA (such as poly-A RNA used as carrier in extractions) was spiked into samples. For the degenerate signatures, only the primer/probe combinations that were able to detect all strains of RVFV tested are listed.