鸭瘟病毒提纯方法比较

Comparison of purification methods of duck plague virus

将鸭瘟病毒CHv株经鸭胚和鸭胚成纤维细胞（DEF）进行培养增殖后，收集病毒液，分别采用沉淀法、层析法、差速离心法和蔗糖不连续密度梯度离心法进行了鸭瘟病毒的提纯研究，结果表明，DEF增殖与蔗糖不连续密度梯度超速离心法是鸭瘟病毒较好的增殖和提纯方法，病毒粒子主要存在于400～500g·L^-1蔗糖层上；经20g·L^-1磷钨酸负染后电镜观察发现，提纯病毒具典型疱疹病毒特征，大小较一致，为170～190nm，由核芯、衣壳和囊膜3部分组成结构较完整的病毒粒子；采用Bradford法测定，纯化病毒液中病毒含量为0．71mg·mL^-1。

Propagated in duck embryo and duck embryo fibroblast monolayer cells （DEF）, CHv strain of duck plague virus was purified respectively by four methods as sedimentation, chromatography, differential centrifugation and sucrose gradient centrifugation. The results indicated there was a best purified effect by sucrose gradient centrifugation from viral culture medium propagated in DEF, purified virions with characteristic of Herpesvirus, which was composed of core, capsid and envelope. The diameter of enveloped virions were 170 - 190nm under transmission electron microscope by using negative staining of 20g·L^-1 phosphotungstic acid, and the banded virus was mainly located between 400 -500g·L^-1 sucrose gradient. And the content of virions in the purified solution was 0. 71 mg·mL^-1 measured by Bradford＇ s method.