Direct Immunofluorescent Assay
F81
cell incubated with CPV-2 in a 96-well plate (Corning, USA) was
prepared. When the CPE was about 80%, the MEM was discarded. Cells were
gently rinsed with PBS three times, then 80% acetone solution was used
to fix cells for 10 min. After washing with PBS three times, CPV-2
fluorescein isothiocyanate (FITC)-conjugated monoclonal antibody (VMRD,
USA) was placed on the wells, which were then incubated in a humid
chamber at 30 °C for 30 min. The stained cells were
observed
with
the
fluorescence microscope (Nikon, Japan).