基于顺磁微粒的金黄色葡萄球菌肠毒素B免疫层析试纸条的制备

Preparation of a rapid paramagnetic particles-based Immumochromatographic test for staphylococcal enterotoxins B

采用基于顺磁微粒标记抗体的免疫层析法,制备金黄色葡萄球菌肠毒素B（SEB）快速定量检测的层析试纸条。利用EDC和NHS交联活化顺磁微粒,对顺磁微粒包被抗体的体系进行优化;根据双抗夹心法原理制备免疫层析试纸条,对制备的SEB免疫层析试纸条进行测试研究。优化的顺磁微粒偶联抗体的缓冲体系为p H 9.0,5mmol/L硼酸,0.05%Tween-20;磁珠包被抗体量为125μg/2mg磁珠,封闭剂为0.25%的脱脂牛奶,磁珠保存液为5mmol/L硼酸、0.05%Tween-20＋0.05%Triton X-100,0.01%Na N3,0.5%BSA,p H9.0;建立的SEB顺磁微粒免疫层析试纸条T线抗体浓度为1mg/m L,最低检测限为1ng/m L,磁信号检测显示,该试纸条在SEB浓度0.1~1 000ng/m L范围内具有很好的线性关系。SEB顺磁微粒免疫层析法具有简单快速、灵敏度高、可定量的特点,可应用于SEB的快速检测。

Magnetic beads were used as labels to establish an immunochromatography test strip for rapid,quantitative of Staphylococcal enterotoxins B（ SEB）. Magnetic beads were catalyzed by EDC / NHS,the coupling system of magnetic beads binding SEB antibody were optimized. An immunoassay test strip in sandwish assay were established,and the detection results of SEB immunoassay test strip were analyzed. 5mmol / L boric acid,p H 9. 0,0. 05% Tween-20 were optimized as the buffer system of magnetic beads binding antibody,125μg antidody were binded to 2mg magnetic beads,0. 25% of skim milk were selected as blocking agent;the prepared magnetic beads were preserved in 5mmol / L boric acid,0. 05% Tween-20 ＋ 0. 05% Triton X-100,0. 01% Na N3,0. 5% BSA,p H9. 0; a magnetic immunoassay strips for detecting SEB was established,the antibody amount immobilized T line was 1mg / m L; the MAR values at 0. 1 ~ 1000 ng / m L SEB presented a better linear correlation,and the detection limit of SEB was 1ng / m L. The SEB immunoassay test strip offers a rapid,high sensitive and quantitative tool for the rapid detection of SEB.