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目的:探索一种新的免疫传感器,以提高检测的敏感性及抗体滴度的线性范围。方法:将碳油墨和银/氯化银油墨滴在PET板,作为两电极测试条,碳为工作电极,用不同的方法把可溶性抗原固定在工作电极的末端,银/氯化银电极作为对照。在电化学工作站中,采用循环伏安法和示差脉冲伏安法测试抗体的效价,并对这些方法的效果进行比较。在此基础上开发2种固定日本血吸虫抗原的印刷电极免疫传感器:戊二醛交联(GA)传感器和壳聚糖戊二醛(Chit-GA)传感器,并采用循环伏安法和示差脉冲伏安法测试抗体滴度检测范围。结果:GA膜修饰电极的日本血吸虫抗原最佳包被浓度为50μg/L,Chit-GA膜修饰电极的最佳浓度值为10μg/L;2种电极的免疫反应时间均为1 min;GA,Chit-GA免疫传感器检测范围分别为1:1000~1:400,1:1000~1:500。示差脉冲伏安法电流的测试值会随着血清效价增大而增大。结论:GA传感器和Chit-GA的交联传感器具有较高的检测灵敏度,反应时间短,检测抗体滴度线性范围宽,并且与日本血吸虫抗体滴度具有良好的线性关系。
Objective: To explore a new immunosensor to improve the sensitivity of detection and the linear range of antibody titers. METHODS: Carbon inks and silver / silver chloride inks were dropped on a PET plate as a two-electrode test strip with carbon as the working electrode. Soluble antigens were immobilized on the working electrode ends using different methods. Silver / silver chloride electrodes were used as controls . Antibodies were tested for potency by cyclic voltammetry and differential pulse voltammetry in electrochemical workstations and the effects of these methods were compared. On the basis of this, we developed two kinds of printed electrode immunosensors immobilized with Schistosoma japonicum antigens: glutaraldehyde (GA) sensor and chitosan glutaraldehyde (Chit-GA) sensor. Cyclic voltammetry and differential pulse voltammetry Antibody test antibody titer detection range. Results: The optimum concentration of Schistosoma japonicum antigen coated on GA membrane electrode was 50μg / L, the optimal concentration of Chit-GA membrane electrode was 10μg / L; the immunoreaction time of both electrodes was 1 min; GA, Chit-GA immunosensor detection range of 1: 1000 ~ 1: 400, 1: 1000 ~ 1: 500 respectively. The value of the differential pulse voltammetry current will increase as the serum titer increases. Conclusion: The cross-linked sensor of GA and Chit-GA has high detection sensitivity, short reaction time, wide linear range of antibody titer, and good linear relationship with the antibody titer of Schistosoma japonicum.