San Gabriel M C, Tohya Y, Sugimura T, Shimizu T, Ishiguro S, Mochizuki M
Laboratories of Veterinary Microbiology, Faculty of Agriculture, Kagoshima University, Japan.
J Vet Med Sci. 1997 Feb;59(2):97-101. doi: 10.1292/jvms.59.97.
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.
在日本分离出的一株犬杯状病毒(CaCV),命名为CaCV No. 48株,在MDCK细胞中增殖,并通过氯化铯平衡梯度离心法纯化。对纯化样品进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析,考马斯亮蓝染色后显示仅存在一种主要的病毒蛋白,约60千道尔顿。使用小鼠超免疫血清进行蛋白质印迹检测到相同的条带,推测为衣壳蛋白条带。这种衣壳蛋白早在接种后2小时就在MDCK细胞中合成。犬的实验性感染导致产生抗CaCV抗体,通过微量中和试验和蛋白质印迹检测到这些抗体。同样,血清学调查不仅显示存在中和抗体,还显示现场血清与纯化病毒的衣壳蛋白有反应性。这些结果表明,CaCV No. 48株的衣壳蛋白具有免疫原性,并且可以在蛋白质印迹中被抗体检测到。