基于重组MTFP基因蛋白的犬心丝虫抗体Dot-ELISA检测方法的建立  

Establishment of Dot-ELISA for Detection on Infection by Dirofilaria immitis in Dogs Based on Recombinant MTFP Protein

作　　者：姜鑫[1,2] 李新[1] 侯洪烈[1,3] 高珺珊[1] 宫鹏涛[1] 李建华[1] 杨举[1] 张西臣[1] 

机构地区：[1]吉林大学动物医学学院,吉林长春130062 [2]黑龙江农业经济职业学院,黑龙江牡丹江157041 [3]辽东学院动物科学系,辽宁丹东118003

出　　处：《天津农业科学》2016年第11期107-110,共4页Tianjin Agricultural Sciences

基　　金：国家公益性行业(农业)科研专项(201303042);辽宁省科学技术计划项目(2012010236-401)

摘　　要：以建立Dot-ELISA检测犬心丝虫血清抗体的方法,用纯化的犬心丝虫MTFP基因蛋白为包被抗原,进行条件优化,筛选最佳反应条件,并进行敏感性、重复性和特异性等试验。结果表明,建立的犬心丝虫Dot-ELISA检测方法最佳抗原包被浓度为0.5μg·片-1;待检样品血清最佳稀释比例为1∶50;HRP标记兔抗犬抗体最佳稀释比例为1∶4 000;建立的Dot-ELISA方法敏感性为1∶200;批间重复试验证明,该方法具有良好的重复性,且与犬等孢球虫阳性血清和犬蛔虫阳性血清没有交叉反应;利用建立的方法对吉林省某地的62份待检血清进行检测,结果阳性率为4.8%。研究成功建立了基于重组MTFP基因蛋白的犬心丝虫Dot-ELISA检测血清抗体方法,为犬心丝虫病流行病学调查和临床诊断提供了新技术。To establish Dot-ELISA for detection infection by Dirofilaria immitis in dogs, the optimum reaction conditions was optimized with purified MTFP protein of Dirofilaria immitis as coating antigen, and the sensitivity, repetition and specificity were tested. The results showed that the optimum reaction conditions of established Dot-ELISA was 0.5μg·tablet-1 for coated concentration, 1：50 for the serum samples and 1：4 000 for rabbit anti canine antibody labeled by HRP. The sensitivity of Dot-ELISA was 1：200. The repeat test showed that the method had a good repeatability. There were no cross reaction with positive serum of Isospora spp and Toxocara canis. The detected results of 62 samples came from Jilin province showed the positive rate was 4.8% by the established method. The Dot-ELISA for detection on infection by Dirofilaria immitis in dogs based on recombinant MTFP protein had successfully estab- lished. It provided a new technology for pidemiological survey and clinical diagnosis of canine dirofilariasis.

关 键 词：犬心丝虫 Dot—ELISA MTFP蛋白 

分 类 号：R383.1[医药卫生—医学寄生虫学]