Multiplication of canine parvovirus in CRFK cells.

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Abstract
Canine parvovirus was inoculated into [Crandell feline kidney] CRFK cells synchronized with thymidine treatment and into the asynchronous cells which were dividing actively. The number of cells bearing intranuclear inclusion bodies was much larger in the former. In the synchronized cells, both infectivity and hemagglutination titers began to rise in the cellular phase 12 h postinoculation and in the extracellular phase 24 h p.i. then reached the maximum 48 and 72 h p.i., respectively. Intranuclear inclusion bodies appeared 10 h p.i., and typical ones with chromatin margination and halo formation were seen 14-16 h. In the immunofluorescence studies, specific fluorescence was detected first in the cytoplasm 4 h p.i. It appeared in the nuclei 6 h p.i. and spread throughout the nuclei 10 h p.i. Its intensity reached the maximum 14-16 h p.i. At 20 h p.i., viral antigen was observed again in the cytoplasm. In the immunoperoxidase studies, development of the stained substance was similar to that in the immunofluorescence method. Immunoperoxidase technique was useful to detect the viral antigen as well as immunofluorescence technique except a weak staining at the early stage of infection. When the virus was inoculated into the synchronized cells 1, 3, 5 or 7 h after the release from thymidine block, the number of cells bearing intranuclear inclusion bodies increased most rapidly in the cells inoculated 3 h after the release, and the fluorescence spread throughout the nuclei 8 h p.i. in the cells inoculated 3 or 5 h after the release.