Objective To isolate CDV and definite diagnosis CD and to compare the sensitivity of vero and Vero.DogSLAMtag (vero-dst) cells in canine distemper virus(CDV).Methods Vero and vero-dst cells were inoculated against standard strain Snyder Hill and clinical accepted CDV positive tissue respectively, after that, comparing them with methods such as CPE, TCID50 and RT-PCR, and then we analyzed their sensitivity to CDV.Results There was no CPE not only after inoculated virus in vero cells but also blind passage for 5 generations, however, within 12h after inoculation, there were caused CPE in vero-dst cells, which was characterized by syncytium formation, and then it was amplified the specific fragment of CDV gene.Conclusions The vero-dst cells have an well-behaved sensitive with CDV, and it is an efficient cell line when isolation of CDV.However, the vero cell line in our lab is not adapt to isolation of CDV.In addition, we isolate a wild strain of CDV from the clinical positive tissue of CD, and we confirm its titer is stronger than standard strain,it can be used for further study.%目的 为了更好地分离犬瘟热病毒(CDV)并确诊犬瘟热,本实验比较了Vero及Vero-dst细胞对此病毒的敏感性.方法 将CDV标准毒株Snyder Hill株及临床犬瘟热阳性犬组织匀浆分别接种Vero及Vero-dst两种细胞,通过观察细胞病变、检测病毒滴度(TCID50),并通过RT-PCR法进行比较,分析两种细胞对CDV的敏感性.结果 接种病毒后Vero细胞盲传5代始终未见细胞病变,而Vero-dst细胞12 h出现了明显的合胞样细胞病变,且RT-PCR扩增出了CDV基因特异性片段.结论 Vero-dst细胞对CDV表现了良好的敏感性,是体外分离培养CDV的一个有效细胞系.而所本实验中使用的Vero细胞并不适于CDV的分离与培养.另外,本实验利用Vero-dst细胞从临床犬瘟热阳性病例中成功分离到了野毒株,并确定其毒力较标准毒株毒力强,可用于进一步的研究.
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