Amperometric immunosensor for detection of antibodies of Salmonella typhi in patient serum
Amperometric immunosensor for detection of antibodies of Salmonella typhi in patient serum
复制标题
DOI：
10.1016/j.aca.2004.10.015
复制
发表时间：
2005-02
影响因子：
6.2
通讯作者：
V. K. Rao;G. P. Rai;G. S. Agarwal;S. Suresh
中科院分区：
文献类型：
--
作者：
V. K. Rao;G. P. Rai;G. S. Agarwal;S. Suresh
开发了一种用于检测患者血清中伤寒沙门氏菌抗体的电流传感器。这涉及使用丝网印刷电极和重组鞭毛蛋白融合蛋白。使用间接酶联免疫吸附测定法检测患者血清中的伤寒沙门氏菌抗体。丝网印刷电极是用聚苯乙烯和石墨制成的。测试了这些电极检测 1-萘酚的能力，1-萘酚是由于碱性磷酸酶水解底物 1-萘基磷酸酯而形成的产物。这些电极涂有通过重组 DNA 技术制成的重组鞭毛蛋白融合蛋白，并用牛血清白蛋白 (BSA) 封闭。此外，将它们与患者血清和山羊抗人碱性磷酸酶缀合物一起孵育。通过使用电流法进行免疫传感。使用来自表面健康个体的混合人类血清样本作为对照。对合并的健康人血清样本和患者血清进行Widal凝集试验和电流分析法。 Widal 测试和电流法之间存在 100% 相关性。电化学法检测所需时间为1小时15分钟，维达尔测试所需时间为18小时。
An amperometric sensor for detection of antibodies to Salmonella typhi in the serum of patients was developed. This involved usage of screen-printed electrodes and recombinant flagellin fusion protein. An indirect enzyme-linked immunosorbant assay was used for detection of antibodies to S. typhi in the patient serum. The screen-printed electrodes were made using polystyrene and graphite. These electrodes were tested for their ability to detect 1-naphthol, which is the product formed due to the hydrolysis of the substrate 1-naphthyl phosphate by the enzyme alkaline phosphatase. These electrodes were coated with recombinant flagellin fusion protein made by recombinant DNA technology and blocked with bovine serum albumin (BSA). Further they were incubated with patient serum and goat anti-human alkaline phosphatase conjugate. The immunosensing was performed by using amperometric method. Pooled human serum samples from apparently healthy individuals were used as control. Both the pooled healthy human serum samples and patient sera were subjected to Widal agglutination test and amperometric method. A 100% correlation was found between the Widal test and amperometric method. The time taken for the detection by electrochemical method is 1h and 15min, while the time taken by Widal test is 18h.