Cg tg k18 ace2 2prlmn j

To check if the DxCell technology can detect active infection in animal samples, we used a mouse model. Briefly, eight (8) heterozygous female K18-hACE c57BL/6J mice [strain 2B6.Cg-Tg(K18-ACE2) 2Prlmn/J; The Jackson Laboratory] were infected intranasally using 50 μL of virus culture (5,000 PFU) per animal, following protocols approved by the Institutional Animal Care and Use Committee at SRI International (#20003), while three (3) mice were not infected (negative controls). Eight days postinfection (DPI-8), the mice were euthanized, and two oropharyngeal swab samples were collected from each animal (51 (link)). The swabs were immediately placed into collection tubes containing 500 μL of complete RPMI medium and transported to the BSL3 lab for processing. For each collected swab, the collection tube was vortexed for 10 to 15 s to release cells into the media before the swab was removed from the collection tubes. The collection tube was then centrifuged at 300 g for 5 min to collect cell pellets. The cell pellet was then resuspended into 50 μL of complete RPMI medium. In a 96-well plate, each sample (cells in 50 μL) was cocultured with 50 μL of 25,000 VHH-Ty1 DxCell in complete RPMI medium. The plate was incubated at 37°C for 24 h before Nluc activity was assessed using the Nano-Glo assay.