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为了寻找一种简单、有效的固定抗体的方法来研制4通道丝网印刷碳电极(4-SPCE)免疫传感器,采用戊二醛交联法将HRP-anti-VP固定于4-SPCE表面,制备了副溶血性弧菌酶免疫电极。通过循环伏安法表征免疫电极和监测酶促反应,根据免疫反应前后还原峰电流下降的百分数(DP)来实现对VP的检测。在优化的免疫反应条件及电化学检测条件下,在103~109cfu/mL浓度范围内,免疫电极免疫反应前后还原峰电流下降的百分数(DP)与VP浓度C的对数呈线性关系,线性回归方程为DP=6.5lgC-3.595,线性相关系数0.9989。该免疫电极具有较好的特异性、重现性(RSD<6%)、稳定性(1周后电流响应为初始值的92%)和准确性(和GB符合率93.3%)。因此,研制出的免疫传感器用于快速筛检VP效果良好。
In order to find out a simple and effective method to immobilize antibody, a 4-channel screen-printed carbon electrode (4-SPCE) immunosensor was prepared. HRP-anti-VP was immobilized on 4-SPCE surface by glutaraldehyde Vibrio parahaemolyticus immune electrode. The cyclic voltammetry was used to characterize the immunodelectrode and to monitor the enzymatic reaction. The detection of VP was based on the percent reduction (DP) of the reduction peak current before and after the immunoreaction. Under the optimized conditions of immunoreaction and electrochemical detection, the percentage of reduction peak current before and after immunoreaction (DP) showed a linear relationship with the logarithm of VP concentration C in the range of 103 ~ 109 cfu / mL. The linear regression The equation is DP = 6.5 lgC-3.595 and the linear correlation coefficient is 0.9989. The immunosensor has good specificity, reproducibility (RSD <6%), stability (92% of initial value after 1 week) and accuracy (93.3% of GB compliance). Therefore, the developed immunosensor for rapid screening of VP works well.