[关键词]
[摘要]
本文研究建立一种检测新型瘦肉精多残留胶体金检测卡的方法,能对大量样品进行定量检测。应用胶体金免疫层析技术,采用柠檬酸钠还原法,标记新型瘦肉精多克隆抗体并喷于玻璃纤维上,新型瘦肉精偶联OVA抗原和羊抗鼠IgG分别结合于硝酸纤维膜上,依次将样品垫、胶体金垫、硝酸纤维素膜和吸水纸组装切割成新型瘦肉精多残留胶体金检测卡。通过灵敏度、假阳性、假阴性试验测试,梯度浓度显色结果表明,该检测卡的灵敏度分别为克伦特罗3 ng/mL、莱克多巴胺5 ng/mL、沙丁胺醇3 ng/mL、西马特罗5 ng/mL、班布特罗5 ng/mL、妥布特罗5 ng/mL、特布他林8 ng/mL、氯丙那林5 ng/mL、马布特罗5 ng/mL、溴布特罗8 ng/mL,检测时间为3 min,检测滴加的最优尿液量为70~90 μL,批内和批间重复性为100%,假阳性率和假阴性率均为0。联检技术能实现现场快速检测,提高检测速度和准确性、降低检测成本,可作为生产过程监管和大批量样本的筛选。
[Key word]
[Abstract]
A new method of multi-residue colloidal gold detection card for detecting lean meat powder was estiblished and could be used for quantitative detection of a large number of samples. Applicated, Be used Colloidal gold immunochromatography assay and sodium citrate reduction method were used to mark the new type lean meat powder antibody, which was then sprayed onto a glass fiber. The new type lean meat powder was coupled with OVA and goat anti-mouse IgG on nitrocellulose membrane, and the sample pad, colloidal gold pad, nitrocellulose membrane and absorbent paper were assembled and cut into multi-mesidue colloidal gold detection cards for new type lean meat powder. The gradient color results obtained by investigating sensitivity, false positiveand false negative, showed that the sensitivities of the detection card were as follows: clenbuterol, 3 ng/mL, ractopamine, 5 ng/mL, salbutamol, 3 ng/mL, sima Tero, 5 ng/mL, bambuterol, 5 ng/mL, tulobuterol, 5 ng/mL, terbutaline, 8 ng/mL, clorprenaline, 5 ng/mL, mabuterol 5 ng/mL, brombuterol 8 ng/mL. The detection time was 3 min, and the optimal urine volume was 70~90 mL. In addition, the intra-and inter-reproducibility was 100%, and the false positive rate and false negative rates were both 0. The joint inspection technology, which could be used to supervise the production process and screen large quantities of samples, could realize rapid detection, improve the detection speed and accuracy and reduce the cost.
[中图分类号]
[基金项目]
科技型中小企业技术创新资金专项项目(2014J4200054)