Detection of Zaire Ebolavirus and Rift Valley Fever Virus in Bat Samples

RNA was extracted from the (1) MARV-experimentally infected bat cage fruit samples, (2) oral swabs collected from the MARV-experimentally infected bats, and (3) rMARV-ZsG-inoculated fruit samples using the MagMAX Pathogen RNA/DNA Kit (Life Technologies, Grand Island, NY, USA) with the MagMAX Express-96 Deep Well Magnetic Particle Processor (Life Technologies, Grand Island, NY, USA) following previously described procedures [26 (link)]. Reverse-transcribed rMARV-ZsG and MARV RNA (all three sample types), RVFV (all sample types except oral swabs), and eukaryotic 18S rRNA (oral swabs only) were detected on the ABI 7500 Real-Time PCR System (Life Technologies, Grand Island, NY, USA) 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 of MARV, the large segment of RVFV, and eukaryotic 18S rRNA gene, respectively (Supplementary Materials Table S1). Relative rMARV-ZsG or MARV log₁₀TCID₅₀ eq/mL were interpolated from a standard curve generated from serial dilutions of the respective virus stocks with known titers in sterile media. It is possible that titers can vary slightly due to the fact that the standard curve was not generated using a saliva matrix.