大片形吸虫免疫胶体金检测试纸条的研制及初步应用

发布时间：2018-06-07 16:57

本文选题：大片形吸虫 + 单克隆抗体　；参考：《中国畜牧兽医》2017年01期

【摘要】：为建立一种快速、简便、适应现场应用的水牛大片形吸虫检测方法,本试验利用抗大片形吸虫ES抗原单克隆抗体杂交瘤细胞株7D1和7D4分别制备和纯化得到单克隆抗体7D1和7D4;采用柠檬酸三钠还原法制备颗粒大小为30nm的胶体金,再用胶体金标记纯化单克隆抗体7D1,在硝酸纤维素膜的质控带和检测带处分别包被羊抗鼠IgG二抗和单克隆抗体7D4,经反应条件优化,组装成了大片形吸虫免疫检测试纸条。经测试该检测试纸条对ES抗原的检测下限为0.94μg/mL,与胰扩盘吸虫、前后盘吸虫、支原体、弓形虫、伊氏锥虫均无交叉反应,特异性好。采集广西地区334份水牛新鲜粪样并用试纸条检测,阳性率为38.62%。结果表明,试纸条方法操作简单、快速、易于判定,特异性好,敏感性高,适合基层大面积普查和现场检测大片形吸虫。
[Abstract]:In order to establish a rapid, simple and suitable method for the detection of buffalo fluke, In this experiment, monoclonal antibodies 7D1 and 7D4 were prepared and purified from hybridoma cell lines 7D1 and 7D4, respectively, and colloidal gold with particle size of 30nm was prepared by sodium citrate reduction method. The monoclonal antibody 7D1 was purified by colloidal gold labeling and coated with sheep anti-mouse IgG second antibody and monoclonal antibody 7D4 respectively at the quality control band and the detection band of the nitrocellulose membrane. The detection limit of es antigen in the test strip was 0.94 渭 g / mL. There was no cross reaction with Pancreatic trematorchiasis, anterior and posterior clonorchiasis, Mycoplasma, Toxoplasma and Trypanosoma evansi, and the specificity was good. 334 fresh faeces of buffalo in Guangxi were collected and tested with test strip. The positive rate was 38.62%. The results showed that the test strip method was simple, rapid, easy to judge, good specificity and high sensitivity. It was suitable for large area survey and field detection of paragonimiasis.
【作者单位】：广西大学动物科学技术学院;河南省动物疫病预防控制中心;
【基金】：广西教育厅科研项目
【分类号】：S855.9

【相似文献】