胶体金免疫层析法对组织样品中磺胺甲噁唑残留的快速检测

Rapid Determination of Sulfamethoxazole Residue by Colloidal Gold Immunochromatographic Method

研究了用于快速检测组织样品中磺胺甲嗯唑残留的胶体金免疫层析检测试剂。采用免疫竞争法，将抗磺胺甲嗯唑多克隆抗体-胶体金复合物包被在胶体金结合垫上，同时将人工合成的磺胺甲嗯唑抗原包被在硝酸纤维素薄膜表面作为检测线（T线），将抗磺胺甲嗯唑多克隆抗体的羊抗兔二抗包被在硝酸纤维素薄膜表面作为质控线（C线）。T线的人工抗原与待测样品中的磺胺甲嗯唑竞争结合胶体金标记的磺胺甲嗯唑多克隆抗体，通过T线与C线的显色对比读出结果。采用该检测试剂检测组织试样时，定量下限可达20μg／L，整个检测过程只需3—5min，且与磺胺嘧啶、磺胺二甲基嘧啶、盐酸克伦特罗、四环素、青霉素、链霉素无交叉反应。检测试剂具有较高的灵敏度及特异性，操作便捷，稳定可靠，可作为组织中磺胺甲嗯唑残留现场监控的有效筛检手段。

A rapid and simple colloidal gold immunochrofnatographic method was established for the determination of sulfamethoxazole（SMZ） residue. Anti-sulfamethoxazole polyclonal antibody labeled by nanocolloidal gold particles was coated in the gold conjugate pad based on the immune competition method： Synthesized sulfamethoxazole conjugated with bovine serum albumin（ SMZ- BSA） was coated on the surface of nitrocellulose filter membrane （NC） as the test line （ T line）, while the anti-anti- body（ goat anti-rabbit IgG antibody, targeting at the sulfamethoxazole polyclonal antibody） was coated on the surface of NC as the control line （ C line）. SMZ - BSA antigen and SMZ combined with SMZ antibody by immune competition reaction. The detection result was obtained by comparing the color of the T line and the C line. The whole detection process could be performed between 3 and 5 min, and the minimum detection concentration could be down to 20 μg/L. No cross-reaction was observed to sulfadiazine, sulfamethazine, clenbuterol, tetracycline, penicillin and streptomycin. The method showed high sensitivity and specification, and could be efficiently used for the in-situ determination of sulfamethoxazole residue in tissue sample.