荧光偏振免疫分析法测定猪肝和鸡肉中的恩诺沙星
Fluorescence Polarization Immunoassay for Determination of Enrofloxacin in Pork Liver and Chicken.
机构信息
Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China.
State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, Wuxi 214122, China.
出版信息
Molecules. 2019 Dec 5;24(24):4462. doi: 10.3390/molecules24244462.
Enrofloxacin (ENR) is a widely used fluoroquinolone (FQ) antibiotic for antibacterial treatment of edible animal. In this study, a rapid and highly specific fluorescence polarization immunoassay (FPIA) was developed for monitoring ENR residues in animal foods. First, ENR was covalently coupled to bovine serum albumin (BSA) to produce specific polyclonal antibodies (pAbs). Three fluorescein-labeled ENR tracers (A, B, and C) with different spacers were synthesized and compared to obtain higher sensitivity. Tracer C with the longest arm showed the best sensitivity among the three tracers. The developed FPIA method showed an IC (50% inhibitory concentration) of 21.49 ng·mL with a dynamic working range (IC-IC) of 4.30-107.46 ng·mL and a limit of detection (LOD, IC) of 1.68 ng·mL. The cross-reactivity (CR) of several structurally related compounds was less than 2%. The recoveries of spiked pork liver and chicken samples varied from 91.3% to 112.9%, and the average coefficients of variation were less than 3.83% and 5.13%, respectively. The immunoassay took only 8 min excluding sample pretreatment. This indicated that the established method had high sensitivity, specificity, and the advantages of simplicity. Therefore, the proposed FPIA provided a useful screening method for the rapid detection of ENR residues in pork liver and chicken.
恩诺沙星（ENR）是一种广泛用于抗菌治疗食用动物的氟喹诺酮（FQ）抗生素。在这项研究中，开发了一种快速且高度特异的荧光偏振免疫分析（FPIA）方法，用于监测动物食品中的恩诺沙星残留。首先，将恩诺沙星与牛血清白蛋白（BSA）共价偶联，以产生特异性的多克隆抗体（pAbs）。合成了三种带有不同间隔臂的荧光素标记的恩诺沙星示踪剂（A、B 和 C），并进行了比较，以获得更高的灵敏度。在这三种示踪剂中，臂最长的示踪剂 C 显示出最佳的灵敏度。所开发的 FPIA 方法的 IC（50%抑制浓度）为 21.49ng·mL，动态工作范围（IC-IC）为 4.30-107.46ng·mL，检测限（IC）为 1.68ng·mL。几种结构相关化合物的交叉反应（CR）小于 2%。猪肝和鸡肉样品中添加的回收率在 91.3%至 112.9%之间，平均变异系数小于 3.83%和 5.13%。免疫分析仅需 8 分钟，不包括样品预处理。这表明所建立的方法具有高灵敏度、特异性和简单的优点。因此，所提出的 FPIA 为快速检测猪肝和鸡肉中的恩诺沙星残留提供了一种有用的筛选方法。
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