羊附红细胞体解离方法的比较研究  

The Comparative Study of Separated Methods of E.ovis

作　　者：韩福生[1] 韩昱[2] 

机构地区：[1]河北北方学院,河北张家口075000 [2]青岛农业大学动物科技学院,山东青岛266109

出　　处：《中国畜牧兽医》2012年第7期249-251,共3页China Animal Husbandry & Veterinary Medicine

基　　金：河北省科技攻关计划项目(07220410)

摘　　要：本试验为建立一种高效的羊附红细胞体解离方法,采集红细胞感染率大于90%的阳性抗凝血,分别应用水浴法和药物体外驱虫法对羊附红细胞体进行解离,观察分离前后附红细胞体感染率、红细胞感染强度、附红细胞体数、杂质含量和附红细胞体的运动性5项指标;提取附红细胞体抗原,制备全蛋白悬液,测定蛋白质含量,比较解离效果。将两种方法制备的羊附红细胞体抗原全蛋白进行SDS-PAGE试验,观察条带是否一致。结果显示,200mL血液中加入1mL双向红莲灭,4℃作用36h,即可达到较好分离效果。与水浴法相比,解离后,红细胞感染率和感染强度明显下降,蛋白含量增加,收率高,纯度好。SDS-PAGE结果显示,体外驱虫法所制的羊附红细胞体抗原全蛋白悬液,其抗原蛋白带与水浴法相同,因此可用于粗制羊附红细胞体抗原。In order to establish an efficient separated method of E. ovis, anticoagulant blood samples whose erythrocyte pos itive infection rates being larger than 90% were collected, the water bath method and drug anthelmintic in vitro method were used to dissociate E. ovis. Eperythrozoon infection rate, erythrocyte infection intensity, Eperythrozoon number, amount of impurity, and Eperythrozoon movement were observed both before and after dissociation. Eperythrozoon antigen was extrac- ted, Eperythrozoon antigen somatic protein suspension was prepared, the protein content was determined, then the dissocia tion effect was compared. SDS-PAGE test on Eperythrozoon antigen somatic protein was used to observe the consistent in band between the two preparations made from different method. The results showed that adding 1 mL ＂Shuang xiang hong lian mie＂ per 200 mL infection blood, reacting 36 h in 4 ℃, could achieve a better separation effect. Comparing with water bath method, after dissociation erythrocyte infection rate and intensity were significantly decreased, the protein content increased （high yield） and got good purity. The SDS-PAGE test results showed that the antigenic protein bands of Eperythrozoon antigen protein suspension from anthelmintic preparation in vitro were same as those from water bath method. The method of drug anthelmin- tic in vitro can be used for the preparation of E. ovis antigen.

关 键 词：羊附红细胞体 解离方法 药物体外驱虫法 SDS-PAGE 

分 类 号：S852.64[农业科学—基础兽医学]