犬细粒棘球绦虫感染胶体金检测试纸条的制备

Preparation of colloidal gold strip for detection of Echinococcus granulosus infection in dogs

目的制备犬细粒棘球绦虫感染胶体金检测试纸条。方法采用柠檬酸三钠还原法制备胶体金颗粒,用于标记抗细粒棘球绦虫Ediag A864单克隆抗体2D12制作金标垫,兔抗细粒棘球绦虫Ediag A864多克隆抗体标记检测线(T线),羊抗鼠Ig标记质控线(C线),制备胶体金试纸条,并进行特异性、敏感性、重复性及稳定性试验。用优化的胶体金检测试纸条对96份犬粪样品(36份参照阳性样,60份临床样品)进行检测。结果制备的犬细粒棘球绦虫感染胶体金检测试纸条与犬蛔虫阳性粪及犬贾第虫阳性粪样品均无交叉反应;粪便稀释度为1∶4时仍可检出正确结果;3次检测参照阳性样本均为阳性;于不同温度(室温、4及37℃)下保存不同时间(1、2、3、4个月)的试纸条均可检出正确结果。96份犬粪便样品中,36份参照阳性样品检出率为97.2%(35/36),60份临床样品均为阴性。结论制备的试纸条具有良好的特异性及稳定性,可应用于临床样品检测及大规模流行病学调查。

Objective To prepare a colloidal gold strip for detection of Echinococcus granulosus infection in dogs.Methods Colloidal gold particles were prepared by sodium citrate reduction method. Colloidal gold strip was prepared by labeling of monoclonal antibody（Mc Ab）2 D12 against Echinococcus granulosus Ediag A864 with colloidal gold particles,while the test（T） line with polyclonal antibody against Echinococcus granulosus Ediag A864,and control（C） line with goat anti-mouse Ig,and tested for specificity,sensitivity,reproducibility and stability. A total of 96（30 positive control and 60 clinical samples） fecal samples of dogs were determined by the developed colloidal gold strip. Results The prepared colloidal gold strip showed no cross reaction with positive fecal samples of Toxocara canis and Giardia canis in dogs. Correct test result was obtained even at a sample dilution of 1 ∶ 4. All the results of positive control in three tests were positive. All the test results of samples stored at various temperatures（room temperature,4 and 37 ℃） for various time periods（1,2,3 and 4 months） were correct. In the 96 samples of dog feces detected by the developed colloidal gold strip,the positive rate of 36 reference samples was 97. 2%（35/36）,while all the results of clinical samples were negative. Conclusion The prepared colloidal gold strip showed high specificity and stability,which might be used for clinical detection and large-scale epidemiological survey of E. granulosus infection.