Viral RNA Extraction and Detection

As previously described14 (link)16 (link), RNA was extracted from gamma-irradiated-Rift Valley fever virus-spiked (RNA extraction-positive control) blood, oral, rectal and urine samples inactivated in lysis buffer solution using the MagMAX Pathogen RNA/DNA Kit (Life Technologies) with the MagMAX Express-96 Deep Well Magnetic Particle Processor (Life Technologies).
Reverse-transcribed MARV and Rift Valley fever virus RNA were detected on the ABI 7500 Real-Time PCR System (Life Sciences) using the SuperScript III Platinum One-Step Q-RT-PCR Kit (Life Technologies), with amplification primers and reporter probes targeting the viral protein 40 gene and the large segment, respectively (Supplementary Table 3). Relative MARV TCID₅₀eq ml⁻¹ were interpolated from standard curves generated from serial dilutions of the titrated MARV 371 bat strain spiked into appropriate biological specimens.