Fluorometric enzyme immunosensing system based on a renewable immunoreaction platform for the detection of Schistosoma japonicum antibody

摘要

A fluoroimmunosensing device which was based on ferulic acid (FA)/horseradish peroxidase system for the detection of Schistosoma japonicum antibody (SjAb) has been developed. To circumvent the difficulty of regeneration of immunocomposite surface, a natural chitosan-epoxy resin matrix was used for the immobilization of SjAg. The surface of the immunocomposite layer reacted was easily regenerated by simple polishing. The renewed surface served as a platform for the competitive immuno-reaction of HRP-SjAb and SjAb with SjAg immobilized at the support body surface and for enzymatic reaction. A novel fluorescent substrate ferulic acid for HRP, which is relatively stable toward H_2O_2, has been adapted in the proposed fluorometric enzyme immunosensing system. FA can been catalyzed to produce a non-fluorescent species. The amount of HRP-SjAb bound to the aforementioned renewable surface layer, which is related to the content of SjAb in samples could be quantitized by measuring the decrease of fluorescence of FA induced by HRP-SjAb. The chitosan incorporated in matrix is favorable for the amplification of this sensing system due to the electrostatic reaction with FA. The proposed method showed a linear response ranging from 45 to 150 ng ml~(-1), with an improved detection limit of 45 ng ml~(-1). The method has been employed to determine SjAb in serum samples.