胶体金免疫层析法检测禽蛋中氯羟吡啶残留

Determination of clopidol residues in poultry eggs by colloidal gold immunochromatography

中文关键词:  氯羟吡啶  胶体金免疫层析法  试纸条  单克隆抗体  禽蛋

英文关键词:clopidol  colloidal gold immunochromatography  test strip  monoclonal antibody  poultry eggs

中文摘要:

      目的 建立胶体金免疫层析法检测禽蛋中氯羟吡啶残留的分析方法。方法 在用间接竞争酶联免疫吸附法分析氯羟吡啶单克隆抗体灵敏度和交叉反应性的基础上, 以胶体金标记单克隆抗体作为示踪标志物, 以包被氯羟吡啶半抗原-卵清蛋白偶联物和羊抗鼠IgG的硝酸纤维素膜作为载体, 制备了氯羟吡啶胶体金免疫层析试纸条。结果 氯羟吡啶单克隆抗体的半抑制浓度(half maximal inhibitory concentration, IC50)为 1.6 μg/L, 与地克珠利、二硝托胺、尼卡巴嗪、磺胺喹噁啉、磺胺二甲嘧啶、癸氧喹酯、盐酸氨丙啉和盐酸氯苯胍8种药物均无交叉反应。制备的胶体金免疫层析试纸条肉眼判定禽蛋中氯羟吡啶的检出限为15 μg/kg, 灵敏度为98%、特异性为93%、假阴性率为2%、假阳性率为7%。结论 制备的试纸条具有操作简单、检测快速、成本低等优点, 可用于大批量禽蛋样本中氯羟吡啶残留的初步筛选。

英文摘要:

      Objective To establish a method for the determination of clopidol residues in poultry eggs by colloidal gold immunochromatography. Methods Based on the analysis of the sensitivity and cross reactivity of clopidol monoclonal antibody by indirect competitive enzyme linked immunosorbent assay, the clopidol colloidal gold immunochromatographic test strip was prepared with the colloidal gold labeled monoclonal antibody as the tracer marker and the nitrocellulose membrane coated with clopidol hapten ovalbumin conjugate and goat anti-mouse IgG as the carrier. Results The half maximal inhibitory concentration (IC50) of clopidol monoclonal antibody was 1.6 μg/L, there was no cross reaction with 8 kinds of drugs, including diclazuril, dinitolmide, nicarbazin, sulfaquinoxaline, sulfadimidine, decoquinate, amprolium hydrochloride and robenidine hydrochloride. The limit of detection of clopidol in poultry eggs was 15 μg/kg, the sensitivity was 98%, the specificity was 93%, the false negative rate was 2%, and the false positive rate was 7%. Conclusion The prepared test strip has the advantages of simple operation, rapid detection and low cost, and can be used for the preliminary screening of clopidol residues in a large number of poultry egg samples.