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羊附红细胞体解离方法的比较研究
引用本文:韩福生,韩昱.羊附红细胞体解离方法的比较研究[J].中国畜牧兽医,2012,39(7):249-251.
作者姓名:韩福生  韩昱
作者单位: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
基金项目:河北省科技攻关计划项目
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,200 mL血液中加入1 mL双向红莲灭,4℃作用36 h,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。

关 键 词:羊附红细胞体  解离方法  药物体外驱虫法  SDS-PAGE  

The Comparative Study of Separated Methods of E.ovis
HAN Fu-sheng , HAN Yu.The Comparative Study of Separated Methods of E.ovis[J].China Animal Husbandry & Veterinary Medicine,2012,39(7):249-251.
Authors:HAN Fu-sheng  HAN Yu
Institution:1. Hebei North University, Zhangjiakou 075000, China;2. College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
Abstract:In order to establish an efficient separated method of E.ovis, anticoagulant blood samples whose erythrocyte positive 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 extracted, Eperythrozoon antigen somatic protein suspension was prepared, the protein content was determined, then the dissociation 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 anthelmintic in vitro can be used for the preparation of E.ovis antigen.
Keywords:E  ovis  separated method  drug anthelmintic in vitro  SDS-PAGE
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