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50% Tissue Culture Infectious Dose assay (TCID50)

JM Julie McAuley
CF Claire Fraser
EP Elena Paraskeva
ET Elizabeth Trajcevska
MS Michelle Sait
NW Nancy Wang
EB Eric Bert
DP Damian Purcell
RS Richard Strugnell
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Tissue culture plates (24 well) were seeded with Vero cells 24 h prior to use and were used at ~ 95% confluency. The samples were serially diluted and inoculated (4 replicates/sample and up to 4 serial dilutions) and incubated for 45 min to enable virus infection of monolayers, then infection media (without FBS) but with TPCK trypsin (1 µg/mL) was added. The plates were incubated (37 °C, 5% CO2) and examined 3 d post-inoculation for virus-induced cytopathic effect (CPE). The TCID50/mL was calculated [5] and plotted as the mean ± standard error of repeated experiments. A two-way ANOVA with Tukey post-tests and p < 0.05 was considered to indicate statistical significance.

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