关键词: 羊附红细胞体; 解离方法; 药物体外驱虫法; SDS-PAGE

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.

Key words: E.ovis; separated method; drug anthelmintic in vitro; SDS-PAGE