Author: Nicola Clementi; Elena Criscuolo; Roberta Antonia Diotti; Roberto Ferrarese; Matteo Castelli; Roberto Burioni; Massimo Clementi; Nicasio Mancini
Title: Combined prophylactic and therapeutic use maximizes hydroxychloroquine anti-SARS-CoV-2 effects in vitro Document date: 2020_3_31
ID: fhy2z49t_6
Snippet: Virus titration. P3 virus stocks were titrated using both Plaque Reduction Assay (PRA, PFU/mL) and Endpoint Dilutions Assay (EDA, TCID50/mL). For PRA, confluent monolayers of Vero E6 cells were infected with 10-fold-dilutions of virus stock. After 1 h of adsorption at 37°C, the cell-free virus was removed. Cells were then incubated for 46 h in DMEM containing 2% FBS and 0.5% agarose. Cells were fixed and stained, and viral plaques were counted. .....
Document: Virus titration. P3 virus stocks were titrated using both Plaque Reduction Assay (PRA, PFU/mL) and Endpoint Dilutions Assay (EDA, TCID50/mL). For PRA, confluent monolayers of Vero E6 cells were infected with 10-fold-dilutions of virus stock. After 1 h of adsorption at 37°C, the cell-free virus was removed. Cells were then incubated for 46 h in DMEM containing 2% FBS and 0.5% agarose. Cells were fixed and stained, and viral plaques were counted. For EDA, Vero E6 cells (4 × 10 5 cells/mL) were seeded into 96 wells plates and infected with base 10 dilutions of virus stock. After 1 h of adsorption at 37°C, the cell-free virus was removed, and complete medium was added to cells. After 48 h, cells were observed to evaluate CPE. TCID50/mL was calculated according to the Reed-Muench method.
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