Viral RNA was extracted with QIAamp Viral RNA Mini Kit (QIAGEN), RNeasy Mini Kit (QIAGEN) and MagMAXTM Viral/Pathogen II Nucleic Acid Isolation Kit (Thermo Fisher Scientific). We quantified viral RNA by real time RT-PCR analysis with a one-step RT-qPCR kit (THUNDERBIRD Probe One-step RT-qPCR kit, TOYOBO) using 5′-ACAGGTACGTTAATAGTTAATAGCGT-3′ for forward primer and 5′-ATATTGCAGCAGTACGCACACA-3′ for reverse primer, and a 5′-FAM-ACACTAGCCATCCTTACTGCGCTTCG-3′ probe, as described (Corman et al., 2020 (link)). The detection limit of SARS-CoV-2 RNA in this study was 39 cycle (Ct value).
Shionoya K., Yamasaki M., Iwanami S., Ito Y., Fukushi S., Ohashi H., Saso W., Tanaka T., Aoki S., Kuramochi K., Iwami S., Takahashi Y., Suzuki T., Muramatsu M., Takeda M., Wakita T, & Watashi K. (2021). Mefloquine, a Potent Anti-severe Acute Respiratory Syndrome-Related Coronavirus 2 (SARS-CoV-2) Drug as an Entry Inhibitor in vitro. Frontiers in Microbiology, 12, 651403.
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