斑点酶联A蛋白免疫吸附试验(Dot-PPA-ELISA)检测猪衣原体抗体方法的建立  

Detection of chlamydial antibody in porcine sera by dot horseradish peroxidase enzyme-linked straphylococcus protein a immnosorbent assay(Dot-PPA-ELISA)

作　　者：张焕容[1] 文心田[2] 曹三杰[2] 

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041 [2]四川农业大学动物科技学院,四川雅安625014

出　　处：《中国预防兽医学报》2007年第8期637-640,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：本研究建立了斑点酶联A蛋白免疫吸附试验(Dot-PPA-ELISA),对猪衣原体IgG的最小检测量为1.21 ng。以间接血凝试验方法为对照,两种方法同时检测120头份猪血清样品,Dot-PPA-ELISA检出43头份(35.83%),效价≥256;间接血凝试验检出26头份(21.67%),效价≥64。诊断膜片与猪瘟、猪布氏杆菌病、猪口蹄疫和猪弓形体病标准阳性血清均不出现有意义的交叉反应。诊断膜片置室温、4℃和-20℃下至少保存3个月其抗原效价不变。试验表明本方法重复符合率为94.44%,且反应效果不受反应板质量和新旧的影响;试剂用量节省,各步骤均可做到严格定量,操作简便,3 h内可出结果,肉眼即可判断,不需要特殊检测仪器。由于辣根过氧化物酶标记的葡萄球菌A蛋白(PPA)是一种广谱诊断试剂,因此Dot-PPA-ELISA还可以用于其它哺乳动物衣原体抗体的检测。A Dot-PPA-ELISA was developed for the detection of chlamydial antibody in porcine sera. A total of 120 swine serum samples were tested. The results showed that Dot-PPA-ELISA detected 35.83 % （43 of 120） positive with an antibody titer of 1：256 or higher. In comparison, Indirect hemagglutination test （IHA） only detected 21.67 % （26 of 120） positive with a titre of 1：64 or higher. The Dot-PPA-ELISA assay was highly specific and did not react with positive sera of swine fever, swine brucellosis, swine foot and mouth disease and swine bow-shaped body disease. It has a high sensitivity of detecting as little as 1.21 ng swine chlamydial IgG. The assay was highly reproducible （94.44 %） and was not affected by the quality of microtiter plate. The chlamydial antigen conjugated NC membranes could be stored at -20 ℃, 4 ℃ or room temperature for at least three months and remain as sensitive. This method was simple, cost effective and could be performed at room temperature in about 3 hours. The Dot-PPA-ELISA could also be used for other mammal because PPA is a broad-spectrum diagnostic reagent.

关 键 词：斑点酶联A蛋白免疫吸附试验 猪衣原体病 检测 抗体 

分 类 号：S852.53[农业科学—基础兽医学]