快速检测食品中四环素类抗生素残留的胶体金免疫层析方法

Colloidal Gold-based Immunochromatographic Assay for Rapid Detection of Tetracycline Antibiotic Residues in Food

摘要:

为了更快速、灵敏、准确快速检测食品中四环素类抗生素残留，该研究利用四环素类（Tetracyclines, TCs）抗生素共同的结构特点制备了识别TCs的广谱单克隆抗体，建立了一种应用于食品中四环素类抗生素残留快速检测的胶体金免疫层析法（Colloidal Gold Immuno-chromatography Assay, GICA）。通过半抗原设计、单克隆抗体制备以及优化抗体标记浓度、人工抗原包被浓度等一系列GICA方法关键因素，实现食品中四环素类抗生素残留的快速检测。结果表明，所制备的广谱单克隆抗体对四环素、金霉素、土霉素、多西环素的半数抑制浓度（IC50）值分别为1.29、1.04、1.34、2.33 μg•L-1。在最优条件下所建立的GICA方法对四环素、金霉素、土霉素检测限为20 μg•kg-1，对多西环素检测限为40 μg•kg-1，检测时间约15 min。该研究建立的GICA方法具有灵敏、准确、快速等特点，可应用于鸡肉、猪肉、草鱼等动物性食品中四环素类抗生素残留的大批量快速筛选。该研究结果为四环素类抗生素广谱单克隆抗体的研究和食品中四环素类抗生素残留检测提供一定理论依据和有效的技术支持。

Abstract:

A rapid, sensitive, and accurate detection method for tetracycline (TC) antibiotic residues in food was developed. Shared structural characteristics of TC antibiotics were exploited for the preparation of a broad-spectrum TC-targeting monoclonal antibody (mAb), and a colloidal gold-based immunochromatographic assay (GICA) was established for rapid detection of TC antibiotic residues in food. Through hapten design, mAb preparation, and optimization of key GICA parameters, such as the antibody labeling and artificial antigen coating concentrations, rapid TC antibiotic residue detection in food was achieved. The half-maximal inhibitory concentrations (IC50) of the broad-spectrum mAb towards TC, chlortetracycline, oxytetracycline, and doxycycline were 1.29, 1.04, 1.34, and 2.33 μg•L-1, respectively. Under optimal conditions, the limit of detection of GICA was 20 μg•kg-1 for TC, chlortetracycline, and oxytetracycline and 40 μg•kg-1 for doxycycline, with a reaction time of approximately 15 min. Therefore, the GICA developed in this study achieved highly sensitive, accurate, and rapid detection, making it suitable for rapid batch screening of TC antibiotic residues in animal food products, such as chicken, pork, and grass carp. The results of this study provide a theoretical basis and effective technical support for future research on broad-spectrum mAbs against TC antibiotics and the detection of TC antibiotic residues in food.