基于阵列倏逝波荧光传感器的雌二醇免疫检测方法

Immunoassay of Estradiol by an Array Evanescent Wave Fluorescent Biosensor

近年来,环境内分泌干扰物愈来愈受到人们的关注。对其用传统的检测技术分析,过程复杂、耗时长、价格昂贵,因此急需发展灵敏度高、反应迅速快、价格低廉的检测手段。介绍了一种平面波导型阵列倏逝波荧光生物传感器,在对系统光路参数进行模拟优化的基础上,可实现24个传感位点的同步、快速、灵敏检测。基于此阵列倏逝波荧光传感器,运用间接竞争免疫检测模式,优化并建立环境内分泌干扰物雌二醇的标准检测条件。实验结果表明,入射角在61.8°时,系统灵敏度最高。该方法对雌二醇检测线性检测区间为0.08~2.52μg·L^(-1),检测限为0.05μg·L^(-1),半抑制浓度为0.46μg·L^(-1),完成单次样品的检测时间为20min。利用该传感器对污水处理厂出水进行加标回收测试,回收率在84%~120%之间,相对标准偏差小于15%,表明该方法能够用于实际水体中雌二醇的快速检测。

In recent years,environmental endocrine disruptors have attracted more and more attention.Traditional detection methods are time-consuming,complex and expensive,which means the development of fast response,high sensitivity and low cost detection method is of vital importance.Herein we described a planar waveguide-based array evanescent wave fluorescent biosensor.Based on the simulation and optimization of the optical parameters of the system,it allows the measurements of up to twenty-four sensing spots simultaneously,rapidly and highly sensitive.By using the indirect competitive immunoassay,the detection conditions of the array evanescent wave fluorescent biosensor were optimized and established for the environmental endocrine disruptor of estradiol.The experimental results show that the system has the highest sensitivity at 61.8°.The linear response ranged from 0.08~2.52μg·L-1 has the detection limit of 0.05μg·L-1 and the 50%inhibition concentration(IC 50)of 0.46μg·L-1.One testcycle was completed within 20 min.The recoveries of estradiol in all sorts ofwastewater effluents ranged from 84%to 120%has relative standard deviation values of less than 15%,confirming the application potential in the measurementof estradiol in reality.