The Chemiluminescent Immunoassay For Triiodothyronine And Zearalenone

The chemiluminescent immunoassay (CLIA) combines the high sensitivity ofchemiluminescent analysis and the specificity of immunoassay, has been widely used in the fieldof biomedicine and environmental science. Based on these points, the dissertation would laystress on the development and application of CLIA methods for determination oftriiodothyronine in human serum and mycotoxin in cereals. This study is showing momentouspractice and academic significance. It consists of five parts:Chapter1In this chapter, the categories, sources and functions of thyroid hormone and thespecies and toxicity of mycotoxin was introduced. On this basis, the analytical methods andtechnologies for triiodothyronine and mycotoxin were reviewed. Also, the basic principles andthe recent study progress of chemiluminescence immunoassay were described and itsdevelopmental prospect was expectedChapter2A solid phase CLIA for the determination of triiodothyronine (T3) in humanserum was established, with the highly sensitive HRP-luminol-H2O2system aschemiluminescence detection system. The rabbit anti-T3antibody was indirectly immobilized onthe96-wells polystyrene microplate to prepare the solid phase antibody. The direct analysis of T3in human serum without extraction was realized in the proposed assay by using8-anilino-1-naphthalenesulfonic acid (ANS) to displace T3from its binding proteins. Comparedwith the commercial chemiluminescent immunoassay, the correlation was good, which provedthe potential clinical application of the method.Chapter3Base on the combination of separation technology of magnetic particle andchemiluminescent immunoassay technology, a magnetic particle-based chemiluminescenceimmunoassay was developed for T3in human serum. The goat anti rabbit antibody which coatedat the surface of magnetic particles, can bond to rabbit anti T3antibody to achieve the separation.This method has been successfully applied to the evaluation of T3in human serum and theresults showed a good correlation with the micropalte chemiluminescent immunoassay.Compared to the micropalte chemiluminescent immunoassay, the proposed method can improvethe detected linear range and sensitivity, needed less amount of antibody, and was easy foroperation and automatization.Chapter4A direct competitive immunoassay for the detection of zearalenone (ZEN) in cereals was established, which based on the HRP-labeled ZEN analog. Magnetic particle was asthe separated solid phase and the HRP-luminol-H2O2system as chemiluminescence detectionsystem. The assay was evaluated and every methodological index can meet the demand ofquantitative analysis. The proposed method has been successfully applied to the evaluation ofZEN in40cereals samples and the results showed a good correlation with the commerciallyavailable ELISA kit.Chapter5The mouse anti-Ochratoxin A (OTA) monoclonal antibody was indirectlyimmobilized on the96-wells polystyrene microplate to prepare the solid phase antibody. TheOTA was labled with HRP, and then a direct competitive immunoassay for the detection of OTAin cereals was established. The proposed method was evaluated and every methodological indexcan meet the demand of quantitative analysis. It can be applicable in the analysis of practicalsamples.