Zeng Shaohua, Tian Zhi, Che Hongli, Yang Huihui, Chen Xiuchun, Feng Qimei, Zhou Yunfei, Zhang Shuju, Wu Zhaoyang, Wang Shiping
Department of Parasitology, Medical College, Hunan Normal University, Changsha 410082, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2012 Jun;37(6):541-8. doi: 10.3969/j.issn.1672-7347.2012.06.001.
To improve the sensitivity and the linear range of electrochemical immunosensor to detect Schistosoma japonicum (S. japonicum) antibody.
Carbon inks and silver/silver chloride inks were printed on a polyethylene terephthalate (PET) board to make a two-electrode test strip, where carbon was the working electrode and S. japonicum soluble egg antigen (SEA) was fixed at one end of working electrode by different methods; silver/ silver chloride electrode was used as control. We tested the valency of the antibody by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in an electrochemistry workstation, and conducted comparison with the results of ELISA. Two new immunosensing electrodes have been developed, based on glutaraldehyde cross-linked (GA) or chitosan-glutaraldehyde cross-linked (Chit-GA) transducer fixing S. japonicum antigen. We tested the titer of the antibody by means of CV and DPV.
Our experimental S. japonicum antigen (50 μg/L) is the optimal test concentration for the GA sensor, and 10 μg/L for Chit-GA sensors. The immune reaction time of both electrodes is all essentially complete in 1 minute. The linear range for S. japonicum antibody in human positive serum sample detection by the glutaraldehyde cross-linked immunosensor is 1:1000 to 1:400, and by the chitosan-glutaraldehyde cross-linked immunosensor is 1:1000 to 1:500. As the concentration of dilution ratio of S. japonicum antibody in human positive serum sample increased, the test value of DPV increased proportionally.
GA sensor and Chit-GA cross-linked S. japonicum sensors have high sensitivity and broad linear range response, and both exhibited a good linear relationship between the DPV signal and the test antibody titer.
提高电化学免疫传感器检测日本血吸虫（日本血吸虫）抗体的灵敏度和线性范围。
将碳墨和氯化银/银墨印刷在聚对苯二甲酸乙二醇酯（PET）板上制成双电极测试条，其中碳为工作电极，日本血吸虫可溶性虫卵抗原（SEA）通过不同方法固定在工作电极一端；氯化银/银电极作为对照。在电化学工作站中通过循环伏安法（CV）和差分脉冲伏安法（DPV）检测抗体效价，并与酶联免疫吸附测定（ELISA）结果进行比较。基于戊二醛交联（GA）或壳聚糖-戊二醛交联（Chit-GA）换能器固定日本血吸虫抗原，开发了两种新型免疫传感电极。通过CV和DPV检测抗体效价。
我们实验中的日本血吸虫抗原（50μg/L）是GA传感器的最佳测试浓度，Chit-GA传感器的最佳测试浓度为10μg/L。两个电极的免疫反应时间在1分钟内基本全部完成。戊二醛交联免疫传感器检测人阳性血清样本中日本血吸虫抗体的线性范围为1:1000至1:400，壳聚糖-戊二醛交联免疫传感器的线性范围为1:1000至1:500。随着人阳性血清样本中日本血吸虫抗体稀释倍数浓度的增加，DPV测试值成比例增加。
GA传感器和Chit-GA交联日本血吸虫传感器具有高灵敏度和宽线性范围响应，并且在DPV信号与测试抗体效价之间均呈现良好的线性关系。
