该研究旨在建立一种简单、快速、高灵敏的鱼、虾和蟹中氯霉素残留的荧光定量免疫层析检测方法,以满足水产品中氯霉素残留检测的需求。通过将氯霉素鼠单克隆抗体G375-5与羧基化铕微球进行偶联标记,氯霉素抗原和羊抗鼠二抗分别包被于硝酸纤维素膜(NC)上作为检测线(T)和控制线(C);根据T线信号值与C线信号值的比值(T/C)和样本中氯霉素浓度建立定量标准曲线;针对鱼、虾、蟹样品,对荧光免疫层析检测方法的灵敏度、准确度和精密度等进行评价,并将其在实际水产样品中与HPLC法进行结果对比。结果表明,该研究建立的时间分辨荧光免疫层析检测方法对水产品中氯霉素的定量限为0.1 μg/kg,添加回收率为73.5%~114.2%,变异系数3.9%~11.5%,与国标仪器方法检测结果的一致性较高。因此,该研究所建立的时间分辨荧光免疫层析检测方法在水产品氯霉素的快速检测工作中具有较高的应用价值和应用前景。
This study aims to establish a simple, rapid and highly sensitive method for the detection of chloramphenicol residue in fish, shrimp and crab, thus to meet the needs of chloramphenicol residue detection in aquatic products. The monoclonal antibody G375-5 to chloramphenicol was labeled with carboxylated europium microspheres, Chloramphenicol antigen and second-antibody were coated on cellulose nitrate membrane (NC) as detection line (T) and control line (C), respectively. The quantitative standard curve was established according to the ratio of T-line signal value to C-line signal value (T/C) as well as the concentration of chloramphenicol in samples. For fish, shrimp and crab samples, the sensitivity, accuracy and precision of fluorescence immunochromatography were evaluated, and the results were compared with those of HPLC in actual aquatic samples. The quantitative limit of chloramphenicol in aquatic products was 0.1 μg/kg. The recovery was 73.5%-114.2%, and the coefficient of variation was 3.9%-11.5%. The results were in high consistency with those obtained by standard instrument. These results showed that the TRFIA method established in this study has high application value and prospects in the rapid detection of chloramphenicol in aquatic products.
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