犬细粒棘球绦虫感染Dot-ELISA检测方法的建立  

Development of Dot-ELISA detection method to Echinococcus granulosus infection in dogs

作　　者：高珺珊 张西臣 刘原源 寇金华 张壮志[2] 李建华 杨举 李赫 李棕松 宫鹏涛 GAO Junshan;ZHANG Xichen;LIU Yuanyuan;KOU Jinhua;ZHANG Zhuangzhi;LI Jianhua;YANG Ju;LI He;LI Zongsong;GONG Pengtao(College of Veterinary Medicine, Jilin University, Changehun 130062, China;Veterinary Research Institute, Xinjiang Academy of Animal Science, Urumqi 830000, China)

机构地区：[1]吉林大学动物医学学院,长春130062 [2]新疆畜牧科学院兽医研究所,乌鲁木齐830000

出　　处：《黑龙江畜牧兽医》2018年第7期105-107,112,249,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家公益性行业(农业)科研专项(201303042);吉林省科技发展计划项目(20130206023NY)

摘　　要：为了建立犬细粒棘球绦虫感染Dot-ELISA检测方法,试验利用抗细粒棘球绦虫EdiagA864兔多克隆抗体进行HRP标记制备酶标抗体,以抗细粒棘球绦虫EdiagA864单克隆抗体2D12作为包被抗体,建立犬细粒棘球绦虫感染Dot-ELISA检测方法,对特异性、敏感性、重复性等进行研究,并用优化的Dot-ELISA检测方法对69份犬粪便样品进行检测。结果表明：所建立的Dot-ELISA检测方法敏感性为1∶8,与犬蛔虫阳性粪便、犬贾第虫阳性粪便均无交叉反应,使用不同批次的膜片均能得到相同结果;应用所建立的Dot-ELISA检测方法对69份犬粪便样品进行检测,其中9份人工感染参照样品的检出率为100%（9/9）,60份长春地区临床样品检测结果为阴性。说明试验建立的犬细粒棘球绦虫感染Dot-ELISA检测方法具有较高的特异性和敏感性,可用于犬细粒棘球绦虫感染的临床检测。The aim of the present study was to establish a Dot-ELISA method for detecting infection of Echinococcus granulosus. Enzyme labeled antibody was prepared by HRP labeling rabbit polyclonal antibody against Echinococcus granulosus EdiagA864. A Dot-ELISA method for the detection of Echinococcus granulosus was established by using anti-Echinococcus granulosus EdiagA864 monoclonal antibody 2D12 as the envelope antibody. The specificity,sensitivity and repeatability of the method were tested. 69 fecal samples of dogs were determined by the developed Dot-ELISA method. The results showed that the sensitivity of developed Dot-ELISA was 1 ∶8. There was no cross reaction with the positive fecal samples of Toxocara canis and Giardia canis. The same results could be obtained by using different batches of diaphragm. The 69 dog feces samples has been detected by Dot-ELISA,in which the detected rates of 9 reference positive samples was 100%（ 9/9） and there were no positive in all of the 60 samples from Changchun. The results indicated that the Dot-ELISA method for the detection of Echinococcus granulosus in this experiment had high specificity and sensitivity and could be used for the clinical detection of Echinococcus granulosus infection.

关 键 词：细粒棘球绦虫 EdiagA864 多克隆抗体 单克隆抗体 DOT-ELISA 检测 

分 类 号：S858.292[农业科学—临床兽医学] S852.734[农业科学—兽医学]