Chemiluminescence enzyme immunoassay (CLEIA) is an analytical method that has extremely high sensitivity, rapidity, simple instrumentation, wide calibration ranges. In this paper, two CLEIA detection methods based HRP-Luminol-H2O2-PIP system were developed respectively, and for their application. The contents of this paper are mainly summarized as follows:1.A competitive cheniluminescence enzyme immunoassay method for β-defensin-2 detection in transgenic mice.This study develops a competitive chemiluminescence enzyme immunoassay (CLEIA) method to determine pBD-2 concentrations in tissue samples of transgenic mice. The linear range of the proposed method for pBD-2 detection under optimal conditions was 0.05-80 ng mL(-1) with a correlation coefficient of 0.9960. Eleven detections of a 30ng mL(-1) pBD-2 standard sample were performed. Reproducible results were obtained with a relative standard deviation of 3.94%. The limit of detection of the method for pBD-2 was 3. pgmL(-1) (3σ). The proposed method was applied to determine pBD-2 levels in the tissues of pBD-2 transgenic mice. CLEIA results showed that the distribution of pBD-2 protein is similar but not identical to that of pBD-2 mRNA, this because the transcription and translation do not always show a simple linear relationship. This finding suggests the importance of pBD-2 analysis at the protein level.2.The construction and application of Spore@Colloidal gold microspheres in the Actinobacillus pleuropneumoniae diagnosis.Suspension arrays of encoded microspheres are emerging as an attractive alternative to traditional enzyme-linked immunosorbent assay(ELISA) and slide-based microarrays. Here, the Bacillus spore was demonstrated as a good candidate of fabricating encoded microparticles to advance immunoassay. The colloidal gold was attached to external surface of spore for the functional microsphere. The Spore@Colloidal gold microspheres were used for immunoassays of animal diseases. The results showed that this Spore@Colloidal gold particle could meet the demand for the daily diagnosis of animal diseases, and has great application and development prospects. |