Vero及Vero-dst细胞对犬瘟热病毒敏感性比较

Comparative Study on the Cell Sensitivity of Canine Distemper Virus in Vero and Vero-dst Cell Lines

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。

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.