Vero E6 cells (ATCC CRL-1586, https://www.atcc.org) and SARS-CoV-2 virus were cultured as described by Heilingloh et al. (7). Neutralization capacity of serum samples was determined by endpoint dilution assay, expressed as 50% tissue culture infective dose (TCID50)/mL. Serial dilutions (1:20–1:2560) of serum samples were incubated with 100 TCID50 of SARS-CoV-2 for 1 h at 37°C and added afterwards to confluent Vero E6 cells cultured in 96-well microtiter plates. On day 3 after infection, the cells were stained with crystal violet (Roth, https://www.carlroth.com) and dissolved in 20% methanol (Merck, https://www.merck.com); we analyzed the appearance of cytopathic effects (CPE) by light microscopy. The neutralizing titer was defined as the reciprocal of the highest serum dilution at which no CPE breakthrough in any of the triplicate cultures was observed.

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