磁分散固相萃取-酶联免疫吸附法检测金黄色葡萄球菌肠毒素A

Detection of staphylococcal enterotoxin A by magnetic-solid phase extraction-enzyme linked immunosorbent assay

中文关键词:  金黄色葡萄球菌肠毒素A  适配体  磁性微球  酶联免疫吸附法

英文关键词:staphylococcal enterotoxin A  aptamers  magnetic microsphere  enzyme linked immunosorbent assay

中文摘要:

      目的 以金黄色葡萄球菌肠毒素A (staphylococcal enterotoxin A, SEA)为代表, 设计并合成一种适配体修饰功能化磁性材料, 与酶联免疫吸附(enzyme linked immunosorbent assay, ELISA)检测技术相结合, 进一步提高检测方法灵敏度。方法 样品经SEA适配体修饰磁性微球富集后, 结合SEA的ELISA试剂盒检测样品中SEA含量, 建立SEA检测的磁分散固相萃取-酶联免疫吸附新方法。结果 与仅采用ELISA试剂盒测定SEA的方法相比, 本研究方法的灵敏度可提高10倍, 检测方法的线性范围为0.01~0.81 μg/L, 同时检测方法具有良好的特异性。将建立的检测方法用于牛奶中SEA的检测, 回收率在84.45%~114.45%之间。结论 本方法操作简单、特异性强、灵敏度高, 同时具有广泛的适用性, 可通过特定适配体修饰磁性微球实现不同目标物的检测, 同时适配体修饰功能化磁性材料也可与其他检测技术联合使用, 提高检测的灵敏度。

英文摘要:

      Objective To design and synthesize an aptamers modified functionalized magnetic material, which is represented by staphylococcal enterotoxin A (SEA) and combined with enzyme linked immunosorbent assay (ELISA) to further improve the sensitivity of the detection method. Methods After the samples were enriched by SEA aptamers modified magnetic microspheres, the SEA content in the samples was detected by ELISA kit using SEA, and a new magnetic dispersion solid-phase extraction-enzyme linked immunosorbent assay method for SEA detection was established. Results Compared with the method only using ELISA kit to determine SEA, the sensitivity of the method in this study was increased by 10 times, and the linear ranges of the detection method were 0.01?0.81 μg/L, meanwhile, the detection method had good specificity. The established method was applied to the detection of SEA in milk with the recovery rates of 84.45%?114.45%. Conclusion This method has the advantages of simple operation, strong specificity, high sensitivity, and wide applicability, the magnetic microspheres modified by specific aptamers can be used to detect different targets, meanwhile, the functionalized magnetic materials modified by aptamers can also be combined with other detection technologies to improve the detection sensitivity.