卡氏住白细胞虫配子体的浓集和纯化

Concentration and Purifcation of Gametocytes of Leucocytozoon caulleayi by Density Gradient Centrifugation

以淋巴细胞分离液和生理盐水为介质 ,分别按不同比例配制成 3种不同密度 (1.0 6 0g/ml、1.0 70g/ml、1.0 73/ml)的分离液 ,并通过无菌采集患卡氏住白细胞虫病的病鸡血液获得了卡氏住白细胞虫的配子体。采用单密度梯度离心法和多密度梯度离心法对该配子体进行了浓集和纯化。进行最佳离心速度和离心时间的筛选结果显示 ,15 0 0r/min离心 2 5分钟分离效果最佳 ;经多密度法离心后可同时获得浓集和纯化的配子体 ;经单密度法离心后 ,只能得到浓集的配子体 ;对浓集的虫体进一步纯化 ,结果显示纯化率可达 95 %以上。通过密度梯度离心的结果判定 ,配子体的密度介于 1.0 70～ 1.0 73g/ml之间。实验结果表明若需要大量分离纯化配子体 ,应以单密度法分二步进行为佳 ;若只需少量纯化的虫体 。

In this study,the gametocytes of Leucocytozoon caulleryi were isolated from blood of diseased chickens by density gradient centrifugation.Using lymphocyte separation solution and physiological saline as separation medium,the two solutions were mixed in different proportion to produce three density separation media.The method of gametocyte separation included the multi-density gradient and the single density gradient centrifugation.The purified and concetrated gametocytes were obtained by the multi-density gradient method and the concentrated gametocytes by the single density gradient centrifugation which were later successfully purified with 1.073g/ml of medium.