2.9. TCID50 Assay

Lung samples were weighed and then homogenized in 1 mL DMEM supplemented with 2% fetal bovine serum, 1 mM L-glutamine, 50 U/mL penicillin, and 50 µg/mL streptomycin with sterile bead. Sample was diluted down a 10-fold gradient and 100 uL of each dilution was transferred to Vero UNC cells. Vero UNC were plated day prior at 10,000 cells/well in a 96-well plate for ~80–90% confluency at time of lung homogenate supernatant transfer. Vero UNC were then incubated with homogenate samples for 6 days before CPE was read. Titers normalized to mg of tissue. TCID₅₀ was calculated using the Reed and Muench method.