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用于快速检测鸡蛋和鸡肉中磺胺类药物的免疫层析侧向流动试纸条的研制。

Development of an immunochromatographic lateral-flow test strip for rapid detection of sulfonamides in eggs and chicken muscles.

作者信息

Wang Xiliang, Li Kui, Shi Deshi, Xiong Ning, Jin Xiue, Yi Jundong, Bi Dingren

机构信息

College of Veterinary Medicine, and National Reference Laboratory of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan 430070, People's Republic of China.

出版信息

J Agric Food Chem. 2007 Mar 21;55(6):2072-8. doi: 10.1021/jf062523h. Epub 2007 Feb 23.

Abstract

A rapid immunochromatographic lateral-flow test strip was developed in the competitive reaction format for the detection of sulfonamides in eggs and chicken muscle. A monoclonal antibody against the common structure of sulfonamides was conjugated to colloidal gold particles as the detection reagent and an N-sulfanilyl-4-aminobenzoic acid (SUL)-bovine serum albumin (BSA) conjugate was immobilized to a nitrocellulose membrane as the capture reagent to prepare the test strip. With this method, it required only 15 min to accomplish the semiquantitative or quantitative detection of sulfonamides. The sensitivity to sulfonamides (sulfamonomethoxine, sulfamethoxydiazine, sulfadimethoxine, and sulfadiazine) was at least 10 ng/mL, as determined with an optical density scanner. By eye measurement, the sensitivity was 20 ng/mL for sulfamonomethoxine, sulfamethoxydiazine, and sulfadimethoxine and 40 ng/mL for sulfadiazine. On the basis of a sulfamonomethoxine standard curve, recoveries were from 89.5 to 95.6% for sulfamonomethoxine, from 89.5 to 95.1% for sulfamethoxydiazine, from 85.0 to 95.6% for sulfadimethoxine, and from 44.8 to 60.9% for sulfadiazine in egg and chicken muscle samples. A parallel analysis of 27 egg samples and 28 chicken muscle samples from the animal experiment showed that the differences between test strips and high-performance liquid chromatography (HPLC) were from 0.8 to 11.2% for egg samples and from 2.2 to 34% for chicken muscle samples for the quantitative detection, and the agreement rates between test strips and HPLC were 100%, based on the maximum allowed residue level of sulfadiazine (100 ng/g) established by the European Union and China. In conclusion, the method is rapid and accurate for the detection of sulfonamides in eggs and chicken muscles.

摘要

研制了一种快速免疫层析侧向流动试纸条,采用竞争反应模式检测鸡蛋和鸡肉中的磺胺类药物。将针对磺胺类药物共同结构的单克隆抗体与胶体金颗粒偶联作为检测试剂,将N-磺胺基-4-氨基苯甲酸(SUL)-牛血清白蛋白(BSA)偶联物固定在硝酸纤维素膜上作为捕获试剂来制备试纸条。用该方法仅需15分钟即可完成磺胺类药物的半定量或定量检测。通过光密度扫描仪测定,对磺胺类药物(磺胺间甲氧嘧啶、磺胺甲氧基嘧啶、磺胺二甲氧嘧啶和磺胺嘧啶)的灵敏度至少为10 ng/mL。通过目视测量,磺胺间甲氧嘧啶、磺胺甲氧基嘧啶和磺胺二甲氧嘧啶的灵敏度为20 ng/mL,磺胺嘧啶的灵敏度为40 ng/mL。根据磺胺间甲氧嘧啶标准曲线,鸡蛋和鸡肉样品中磺胺间甲氧嘧啶的回收率为89.5%至95.6%,磺胺甲氧基嘧啶为89.5%至95.1%,磺胺二甲氧嘧啶为85.0%至95.6%,磺胺嘧啶为44.8%至60.9%。对动物实验中的27份鸡蛋样品和28份鸡肉样品进行平行分析表明,在定量检测中,试纸条与高效液相色谱法(HPLC)之间的差异,鸡蛋样品为0.8%至11.2%,鸡肉样品为2.2%至34%;基于欧盟和中国规定的磺胺嘧啶最大残留限量(100 ng/g),试纸条与HPLC的符合率为100%。总之,该方法用于检测鸡蛋和鸡肉中的磺胺类药物快速且准确。

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