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with serial 10-fold dilution of virus.
SF767 cells (obtained from J. Loftus) grown to 50% confluence were incubated for 24 hours in a 1∶1 dilution of virus:media with 4 µg/ml Polybrene.
In the parallel SF767 and MCF7 infection experiment, a 1∶4 dilution of virus:media with 6 µg/ml Polybrene was used.
The fifty percentage tissue culture infection dose (TCID50) of virus stocks was determined by infecting MDCK cells with serial 10-fold dilution of virus and calculated by the method of Reed and Muench [12].
The Tissue Culture Infectious Dose (TCID50) was determined for each viral stock per 1 ml by identifying the dilution of virus in which 50% of the TZM-bl cells were infected, as measured by the presence of β-galactosidase.
To determine 50% mouse lethal dose (MLD50) of the HPAI H5N1 virus A/Shenzhen/406H/06, groups of 5 mice were inoculated i. n. with serial 10-fold dilution of virus.
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The tissue sections also harboured unaffected tissue, leading to the dilution of virus-containing cells.
The sensitivity was determined by testing serial dilutions of virus and RNA standard.
In the three dilutions of virus analyzed, viral titers increased thereafter until animal death.
Serial dilutions of virus were prepared in 96 well plates using medium containing 75 µg/ml of DEAE-dextran.
For plaque assay, serial 10-fold dilutions of virus sample were added into a monolayer of MDCK cells.
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