Identification of canine calicivirus capsid protein and its immunoreactivity in western blotting.
A canine calicivirus (CaCV) isolated in Japan, designated as CaCV No. 48 strain, was propagated in MDCK cells and purified by CsCl equilibrium gradient centrifugation. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of the purified samples revealed the presence of only one major species of viral protein of about 60 kilodaltons after Coomassie staining. The same band, presumably that of the capsid protein, was detected by western blotting using a mouse hyperimmune serum. This capsid protein was synthesized in MDCK cells as early as 2 hr post-inoculation. Experimental infection of dogs resulted in the production of anti-CaCV antibodies which were detected by microneutralization test and western blotting. Likewise, serosurvey revealed not only the presence of neutralizing antibodies but also reactivity of the field sera against the capsid protein of the purified virus. These results indicate that the capsid protein of CaCV No. 48 strain is immunogenic and could be detected by antibodies in western blotting.
San Gabriel
M C
MC
Laboratories of Veterinary Microbiology, Faculty of Agriculture, Kagoshima University, Japan.
Tohya
Y
Y
Sugimura
T
T
Shimizu
T
T
Ishiguro
S
S
Mochizuki
M
M
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015153:Blotting, Western / Q000379:methods / Q000662:veterinary; D002139:Caliciviridae / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D017250:Caliciviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D002213:Capsid / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D004283:Dog Diseases / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000662:veterinary; D007668:Kidney / Q000473:pathology / Q000821:virology; D051379:Mice; D013047:Specific Pathogen-Free Organisms; D013997:Time Factors
D000914:Antibodies, Viral