摘要
摘要:为了探索压阻式微悬臂梁免疫传感器用于小分子T-2毒素的检测效果及其动力学响应规律.基于生物素一亲和素放大系统构建了一种操作简单、快速灵敏的非标记传感器。该传感器对T-2毒素的检测限为4txg/L(S/N≥3),检测的线性范围为4—60μg,L,线性回归方程为AUe=0.447C+1.315(n=5,R=0.983,p〈0.001)。传感器具有很好的特异性和重现性,能满足河水、土壤、食品等实际模拟样品的检测要求。基于配体一受体动力学和吸附动力学创建了两种压阻式微悬臂梁免疫传感器检测小分子T-2毒素的响应动力学模型.对T-2毒素实际检测数据进行拟合分析。结果表明:创建的这两种模型均能较好地拟合T-2毒素实际检测数据,相关系数R值在0.9689以上(p〈0.001)。基于配体一受体特异结合动力学模型由于考虑了敏感膜与靶分子间的特异相互作用,对T-2毒素实际检测数据的拟合效果更佳。相关系数R值在0.9703以上(p〈0.001),能更好的反映压阻式微悬臂梁免疫传感器检测小分子T-2毒素的响应特点和规律;并且能求出更有意义的平衡响应电压(△Ue)、响应时间(tO)等参数.且根据曲线拟舍方程求出的△Ue、t0均与实测值非常接近.
In order to evaluate the test performance and investigate the response dynamic characteristics of small- molecule T-2 toxin detection by piezoresistive microcantilever immunosensor, a simple, rapid, sensitive and non-labeled sensor was proposed based on biotin-avidin binding system. Results showed that the detection limit of T-2 toxin was 47g L? 1 (S/N ≥ 3). A linear relationship between the equilibrium response voltage and the concentration of T-2 toxin in the range of 4?60μg/L was obtained, with the linear regression equation of AUe=0.447C+l.315 (n=5, R=0.983, p〈0.001). The sensor indicated a good specificity and reproducibility, which could meet the requirements in determination of simu- late samples, such as river water, soil and foodstuff sample. Two reaction kinetic models of small-molecule T-2 toxin detection by piezoresistive microeantilever immunosensor based on receptor-ligand kinetics and adsorption kinetics were es- tablished, respectively. The built models were used to fit the measured data of T-2 toxin. Results showed that both these two models could well fit the measured data of T-2 toxin, the correlation coefficient R was greater than or equal to 0.9689(p〈0.01 ). The model based on specific binding of ligand-receptor could fit the measured data better since it con- sidered the specific interaction between sensitive molecules and target molecules. The correlation coefficient R was greater than or equal to 0.9703 (p〈0.01), which could better reflect the response kinetics of small-molecule T-2 toxin detection by piezoresistive microcantilever immunosensor. And it could give some more meaningful parameters, such as equilibrium response voltage (?Ue) and response time (tO). Equilibrium response voltage (?Ue) and response time (tO) ob- tained from the fitting equation fitted well with the measured values.