检测多种动物弓形虫抗体的SPA-ELISA方法的建立及初步应用  

Development and application of SPA-ELISA for detection of anti-Toxoplasma antibodies in sera from many species of animals

作　　者：张东林[1,2] 惠煜[1,3] 周艳琴[2] 董海岚[1,3] 方瑞[1,2] 聂浩[1,2] 王正松[1,2] 赵俊龙[1,2] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,湖北武汉430070 [2]华中农业大学动物医学院,湖北武汉430070 [3]河南省南阳市兽医站,河南南阳473010

出　　处：《中国兽医学报》2009年第9期1163-1166,共4页Chinese Journal of Veterinary Science

基　　金：湖北省科技攻关计划资助项目(2006AA205A02;2007AA201AA);教育部重点科研资助项目(105120)

摘　　要：利用重组弓形虫MIC3作为包被抗原，SPA作为偶联物建立了可检测多种动物弓形虫抗体的SPA—ELISA方法。确定了抗原最适包被质量浓度为2．6mg／L；血清最适稀释度为1：160，作用时间为45min；优化了酶标SPA工作浓度，作用时间及底物显色时间。对已知阳性血清的检测下限可达1：6400，其敏感性是IHAT方法的50～100倍。对临床4种动物共332份血清进行检测，阳性率由高到低依次为：SPA-ELISA（39．4％）〉MAT（39．1％）〉IHAT（13．1％）。以上结果表明，SPA-ELISA不失为一种适用于多种动物弓形虫抗体检测的理想方法。In order to detect Toxoplasma gondii infection in many species of animals, an ELISA using SPA as conjugate （SPA-ELISA） was developed and used to detect anti-T, gondii antibodies in clinical samples of pigs, goats,dogs and cats. The assay was optimized for an antigen coating concentration of 2.6 mg/L and a serum dilution of 1 ： 160,with a standard incubation time of 45 min. The control positive serum could be detected at a maximum dilution of 1 ： 6 400. The 332 clinical serum samples from four species of animals were tested,and the detection methods ranked as follows in descending order of sensitivity： SPA-ELISA（ 39.4 % ）〉 MAT（ 39. 1% ）〉IHAT（ 13. 1% ）. All these results indicated that SPA-ELISA could be a good method for serological detection of T. gondii antibodies in many species of animals.

关 键 词：弓形虫 重组MIC3 SPA—ELISA 

分 类 号：S852.7[农业科学—基础兽医学] R535[农业科学—兽医学]