庆大霉素快速Dot-ELISA检测方法的建立及初步应用

Development and Application of Dot-ELISA for Rapid Detection of Gentamicin Residue in Milk

本研究旨在建立一种适用于快速、现场检测要求的庆大霉素残留Dot-ELISA检测方法。将硝酸纤维素膜用激光切割成小圆片,粘贴到激光打孔的塑料胶条上,制成8个圆片组成的NC膜检测条。通过条件优化,建立了基于NC膜条的庆大霉素残留直接竞争Dot-ELISA检测方法。结果表明,该方法可以快速检测牛奶等食品中的庆大霉素残留,检测灵敏度达到60 ng/mL,可用于庆大霉素残留的筛查。与微孔板ELISA相比,Dot-ELISA可以大大节省检测时间。直接竞争Dot-ELISA具有简便、快速、灵敏、直观的特点,具有推广和应用价值。

A dot enzyme linked immunosorbent assay（Dot-ELISA） was established for rapid detection of gentamicin residue in milk on-site.The Dot-ELISA was performed on modified nitrocellulose membrane strip（NCM strip） which had eight round pieces of nitrocellulose membrane with diameter of 2 mm,which was cut by Versa Laser,and then was affixed on a plastic adhesive tape.Based on the NCM strip,a direct competitive Dot-ELISA was developed for detecting gentamicin and the influence of several factors on the immunoassay was studied.The optimized Dot-ELISA has been used to detect gentamicin in milk and a limit detection of 60 ng/mL was obtained.Compared with the normal ELISA tests carried on the microtiter plates,the detection time was saved evidently.The Dot-ELISA was characterized by easy use,rapid reaction,economical feasibility,and observable results and the method was worth promoted.