建立氯霉素,克伦特罗和雌二醇3种兽药残留同时检测的悬浮芯片法。通过将3种兽药的BSA蛋白结合物偶联于悬浮芯片的固相载体——聚苯乙烯荧光微球上作为检测探针,采用间接竞争法,在液相反应体系中,3种小分子兽药抗原和微球上的兽药结合物共同竞争液相中各自特异性的生物素化单抗,再加入藻红蛋白标记的链霉亲和素,反应后检测获得荧光信号,绘制出3种兽药残留检测的标准曲线。同时进行3种兽药的常规酶联免疫吸附法标准曲线的测定。在检测技术、检出限、检测区间、特异性、盲样测定和多元分析等方面对两种方法进行比较。除了特异性外的其它指标的比较中,悬浮芯片法均具有明显优势。两种方法的特异性检测具有良好的一致性。高通量悬浮芯片技术,具有操作简单、灵敏快速和成本低廉等优点,为多种兽药残留的快速检测提供了新方法。
A novel suspension array technology was established for the detection of three kinds of veterinary drug residues: chloramphenicol, clenbuterol and 17-β-estradiol. The three conjugates in which veterinary drugs coupled with BSA were immobilized on the solid carrier of the suspension microarray-polystyrene fluorescent microspheres/beads as detective probes. Indirect competitive technology was employed. Competitive reac- tions between the veterinary drugs in the aqueous phase and the veterinary drugs-BSA conjugates on the beads for coupling with their complimentary specific biotinylated monoclonal antibodies were carried out. And then, straptavidin-phycoerythrin was added for coupling and the fluorescent signals were captured. Afterwards the detective standard curves were plotted. The regular ELISA standard curves of the three veterinary drugs were also plotted. Comparison between suspension array and regular enzyme-linked immunosorbent assay (ELISA) was in the respects of the detective technology, the detection limits, the detective ranges, the samples detection and the multi-analysis. Suspension array technology is distinct advantageous except for specificity. There was well consistent performance between the two methods. The high-throughput suspension array provides a novel method for multi-analysis of veterinary drugs with simple operation, sensitive, rapid and low costing.