一种培养鉴定风疹病毒的简便试验方法的建立  

A simple method for isolation and detection of rubella Virus in Vero cells

作　　者：方草晖[1] 胡勇[1] 张文艳[1] 类延花[1] 金晶[1] 吴建军[1] 王明丽[1] 

机构地区：[1]安徽医科大学微生物教研室,合肥230032

出　　处：《安徽医科大学学报》2004年第3期241-243,共3页Acta Universitatis Medicinalis Anhui

基　　金：国家教育部科学技术研究重点项目 (编号 :0 10 5 2 );安徽省生物医药重大科技专项项目 (编号 :0 13 0 3 0 0 3 )

摘　　要：目的　建立使风疹病毒 (RV)在Vero细胞 (非洲绿猴肾传代细胞株 )上产生的细胞病变效应 (CPE)易于观察的简便试验方法 ,以利于病毒的分离、培养与鉴定。方法　在RV接种Vero细胞 4天后 ,用不同 pH值 (4 0、4 5、5 0、5 5、6 0、7 0 )的融合营养液处理细胞 ,继续按常规条件培养 1天后 ,在倒置显微镜下观察RV引起的特征性CPE 多核巨细胞 ,染色后计数多核巨细胞的形成率。同时检测在不同条件(融合时间、病毒接种初始滴度 )下 ,多核巨细胞形成率的变化。结果　感染RV的Vero细胞经过酸性融合营养液的处理 ,可出现RV特征性CPE 多核巨细胞。发现在 37℃、pH5 0的酸性融合营养液处理染毒细胞 10min后 ,继续培养 1天 ,出现的细胞病变形态最典型 ,染色后计数发现多核巨细胞的形成率最高 ,为 14 7% (P <0 0 1)。融合处理的时间对多核巨细胞的形成率没有影响。随着病毒接种初始滴度的降低 ,多核巨细胞的形成率也相应降低。结论　此方法简单、易行 ,使RV在Vero细胞上产生的细胞病变清晰易观察。Objective To establish a method which is used to facilitate the isolation, detection and culture of Rubella Virus (RV) in Vero cells, making the cytopathic effect (CPE) caused by RV easy to recognize. Methods After 4 days post infection, Vero cells were treated with acid fusion medium. Then the cells were cultured in neutral medium for 1 day and stained. The specific CPE caused by RV-gigantic cells with multiple nuclei were observed. The effect of fusion treatment time and virus infectivity titer on polykaryocyte prduction were also tested. Results Vero cells which are treated for 10 min with acid fusion medium in pH 5.0 displayed typical gigantic cells with multiple nuclei, while control groups had no change. Variation of the reaction time of the fusion treatment had no effect on the extent of cell fusion. The percentage of fused cells decreased while the virus infectivity titer decreased. Conclusion This method is simple and accessible, making the CPE of RV in Vero cells easier to recognize than that of normal method.

关 键 词：风疹病毒 致细胞病变 病毒 巨细胞 

分 类 号：R373.11[医药卫生—病原生物学] R373.9[医药卫生—基础医学]