Development Of Enzyme-Linked Immunosorbent Assay Test Kit For Sulfamethoxydiazine Residues

Sulfonamides are synthetic bacteriostatic drug, widely used in animal husbandry and as feed additives. Food derived from animals treated with sulfonamides may be contaminated with those drugs, and could result in resistance in pathogenic organisms because of the extensive use of sulfonamides, and will influence human health.To safeguard the public heath, some countries have established total sulphonamides tolerance limit in tissues for human consumption at 0.1mg/kg or even none. Many chemical and physics methods are used to be detecting sulfonamides. Chromatographic analysis and Enzyme-linked immunosorbent assay(ELISA) method are often applied in drug residues assay.Though some of these chromatographic analysis methods are sensitive and accurate, they are not suitable for screening a large number of samples, for they are complex of pre-treatment, time- consumption, costly, needing high skill and valuable equipment. Enzyme-linked immunosorbent assay (ELISA) as a rapid, special, sensitive and high-performance biological method is gradually popular in drug residues assay. Developing a fast ELISA detection kit can be expediently applied in practice. Immunoassay will be the main detecting methods in drug residues and be the developing direction.This research synthesized the coating antigen and devoloped the antibodies, and established the ELISA method for detecting SMD. In the experimental condition, the residues of SMD in chook's tissues were determined.We resuscitated the cell lines of monoclonal antibodies, and cloned the positive cells by limiting dilutions. This procedure resulted in six stable clones: B6, D12, E2, H5, G8 and G11. Mice were injected intraperitoneally with H5 cells to produce ascites.We do a chessboard test to confirm the work concentration between producing ascites and antigen. The results are: the work concentration of antigen is 1:5000, and the producing ascites is 1:100000.We make an experiment on chook that was given some SMD. The samples of muscle, liver and kidney were collected. We perfected the pre-treatment of samples and...