The Application Of HRP-Luminol-H₂O₂-PIP Chemiluminescence System In The Enzyme Immunoassay

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.