鸡、鸭肉中金刚烷胺、金刚乙胺、索金刚胺间接竞争ELISA检测方法研究

目的:建立一种检测鸡肉、鸭肉中金刚烷胺、金刚乙胺、索金刚胺的间接竞争酶联免疫吸附方法（Indirect competitive enzyme-linked immunosorbent assay，ic-ELISA）。方法:本研究基于间接竞争酶联免疫方法的原理，在酶标板微孔中预包被偶联抗原，样本中含有的金刚烷胺、金刚乙胺、索金刚胺与微孔中预包被的偶联抗原特异性地竞争酶标记抗体，催化底物显色，根据显色深浅来计算样本中金刚烷胺类药物的含量。结果:金刚烷胺、金刚乙胺、索金刚胺的检测限分别为:0.57、0.42、0.41 μg/kg （鸡肉）和0.59、0.40、0.38 μg/kg （鸭肉）；定量限分别为0.85、0.63、0.69 μg/kg （鸡肉）和0.94、0.68、0.52 μg/kg （鸭肉）；添加回收率范围为67.0%~117.9%；日内变异系数6.3%~12.7%，日间变异系数8.1%~14.5%，且实际样品检测结果与HPLC-MS一致性较高（R2=0.9990），表明该方法具有良好的准确度和精密度。结论:本研究建立的ic-ELISA方法适用于鸡鸭肉样品中金刚烷胺类药物残留的检测，方法的灵敏度高、稳定性好，可应用于批量样本的快速筛查，具有良好的实际应用价值。

Indirect Competitive ELISA for Detection of Amantadine,Rimantadine and Soramantadine in Chicken and Duck Meat

Objective: To establish an ic-ELISA for the determination of amantadine,rimantadine and soramantadine in chicken and duck meat. Methods: Based on the principle of ic-ELISA,conjugated antigen was pre-coated in the wells of microtiter plate. Amantadine,rimantadine,and soramantadine contained in the samples will competitively combine with the enzyme-labelled antibody,which catalyzing colour reactions of the substrates. The concentration of amantadines in the sample was detremined on the basis of OD value. Results: The detection limits of amantadine,rimantadine and somantine were respectively 0.57,0.42,0.41 μg/kg(chicken),and 0.59,0.40,0.38 μg/kg(duck). The limits of quantification were respectively 0.85,0.63,0.69 μg/kg(chicken),and 0.94,0.68,0.52 μg/kg(duck). The added recovery range was 67.0%~117.9%;the intra-day and inter-day variation were 6.3%~12.7% and 8.1%~14.5%,respectively. The actual sample detection results were well consistent with HPLC-MS(R2=0.9990),showing that the method had good accuracy and precision. Conclusion: The ic-ELISA method established in this study is suitable for the detection of amantadine residues in chicken and duck meat samples. The method has high sensitivity and good stability. It can be used for rapid screening of batch samples and has good practical applications value.