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MDCK cells were seeded on 96-well tissue culture plates 1 day before the viral titration assay. Cells were washed once with PBS and changed to serum-free MEM medium with 1% PS and L-1-Tosylamide-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin. Virus samples or culture supernatants were titrated in serial half-log10 dilutions with serum-free medium prior to the addition of the diluted virus to MDCK cell plates in quadruplicate. The highest viral dilution leading to cytopathic effect (CPE) in ~ 50% of inoculated wells was estimated using the Karber method.
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