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Comparison of Subgenomic and Total RNA in SARS-CoV-2-Challenged Rhesus Macaques

2021; American Society for Microbiology; Volume: 95; Issue: 8 Linguagem: Inglês

10.1128/jvi.02370-20

1098-5514

Gabriel Dagotto, Noe B. Mercado, David R. Martinez, Yixuan J. Hou, Joseph P. Nkolola, Robert H. Carnahan, James E. Crowe, Ralph S. Baric, Dan H. Barouch,

Animal Virus Infections Studies

Respiratory virus challenge studies involve administration of the challenge virus and sampling to assess for protection from the same anatomical locations. It can therefore be difficult to differentiate actively replicating virus from input challenge virus. For SARS-CoV-2, specific monitoring of actively replicating virus is critical to investigate the protective and therapeutic efficacy of vaccines, monoclonal antibodies, and antiviral drugs. We developed a SARS-CoV-2 subgenomic RNA (sgRNA) RT-PCR assay to differentiate productive infection from inactivated or neutralized virus. Subgenomic RNAs are generated after cell entry and are poorly incorporate into mature virions, and thus may provide a marker for actively replicating virus. We show envelope (E) sgRNA was degraded by RNase in infected cell lysates, while genomic RNA (gRNA) was protected, presumably due to packaging into virions. To investigate the capacity of the sgRNA assay to distinguish input challenge virus from actively replicating virus