刚地弓形虫RH株SAG3蛋白抗体检测的ELISA方法建立  

Establishment of ELISA for Detecting Protein SAG3 Antibody of RH Strain of Toxoplasma gondii

作　　者：马亮[1,2] 任保彦[1] 左绍志[1] 宫鹏涛[1] 李建华[1] 张国才[1] 杨举[1] 张西臣[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]吉林省吉林市万达兽药经销公司,吉林吉林132001

出　　处：《安徽农业科学》2012年第15期8527-8528,8550,共3页Journal of Anhui Agricultural Sciences

基　　金：国家科技支撑计划课题(2008BAD96B11-3)

摘　　要：[目的]利用纯化的刚地弓形虫RH株表面蛋白SAG3的原核重组蛋白为基础,建立抗体间接ELISA检测方法。[方法]采用棋盘滴定法,确定间接ELISA的最佳条件。采用鼠抗新孢子虫阳性血清、鼠抗安氏隐孢子虫阳性血清、鼠抗柔嫩艾美尔球虫阳性血清、鼠抗蓝氏贾第虫阳性血清以及鼠抗微小隐孢子虫阳性血清和健康小鼠阴性血清作对照,检测血清之间是否有交叉反应。[结果]最佳包被条件为:抗原包被浓度0.25μg/孔,被检血清稀释度1∶400,酶标二抗稀释度1∶3 000,封闭剂5%脱脂奶粉的PBST溶液;血清之间没有交叉反应。该方法的灵敏度为1∶1 600。[结论]该方法灵敏度高,特异性较强,可重复性较好,可用于对弓形虫感染的检测及流行病学调查。[ Objective ] The research aimed to establish the indirect ELISA for detecting the purified surface protein SAG3 antibody of Toxoplasma gondii（ T. gondii）. [ Method ] Using chessboard titration method, the best conditions of indirect ELISA were determined. Taking negative serum as control, the positive serums of mouse against Neospora caninum, G. lamblia, C. andersoni, E. tenella and C. parvum were used to detect if the indirect ELISA has cross reactions with them. [ Result ] The optimal coating conditions were as follows：antigen coating concentration was 0.25 ug/hole, and the optimal dilution of the detected serum of T. gondii and enzyme-labeled second antibody were 1 ： 400 and 1：3 000 respectively, blocking agent was the PBST solution of 5% honfat-dry milk. The developed indirect ELISA had no cross reaction with positive serums. The sensitivity of this method was 1：1 600. [ Conclusion ] This method had high sentivily, stronger specificity and better repeatability, which might be used for the diagnosis and epidemiological investigation of T. gondii with SAG3 protein.

关 键 词：刚地弓形虫 SAG3 表面蛋白 间接ELISA 

分 类 号：S851.347.103[农业科学—预防兽医学]