牛奶中β-内酰胺酶抑制剂——舒巴坦直接竞争ELISA的分析研究

Research on the direct competition ELISA analysis ofβ-lactamase inhibitor-sulbactam in milk

建立代表性的β-内酰胺酶抑制剂-舒巴坦直接竞争酶联免疫吸附分析法(dc ELISA)。通过对封闭液浓度、离子浓度、p H、包被条件等分析条件优化后建立dc ELISA,并进行应用。结果表明方法的IC_(50)为5.25 ng/m L,IC_(15)为1.07×10^(-3)ng/m L,板内和板间平均变异系数分别为6.69%、9.02%,对他唑巴坦和克拉维酸的交叉反应率分别是7.12%、0.93%。牛奶样品的添加回收率在85.66%~89.23%之间。该方法灵敏度较高,稳定性较好,特异性较强,可以应用于乳制品中舒巴坦残留的快速检测。

The aim of this study was to establish a direct competitive enzyme - linked immunoassay （ dc - ELISA ） method to detect the representative β-lactamase inhibitor-sulbactam in dairy products.After analyzing the condition of blocking solution concentration,ion concentration, coating conditions, pH, and other optimization, this developed method presented an IC50 of 5.25 ng/mL,a detection limit IC15 of 1.07 × 10-3 ng/mL. Intra- and inter-batch relative standard deviations（RSD） were 6.69% ,9.02% ,respectively.The cross-reactivity rates for tazobactam and clavulanic acid were 7.12% ,0.93%, respectively. The recoveries of milk in spiked samples ranged from 85.66% -89.23%.The developed dcELISA method was satisfied for the the field of mass rapid screen of sulbactam residue in milk samples.