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Study on Colloidal Gold Immunochromatography Technique for Rapid Detection of Chloramphenicol Residues in Food

Author: XuHuan
Tutor: TianSuJuan
School: Guangdong College of Pharmacy
Course: Of Pharmacy
Keywords: Colloidal gold Chloromycetin Immunochromatographic Monoclonal antibodies Rapid detection
CLC: R155.5
Type: Master's thesis
Year: 2010
Downloads: 151
Quote: 4
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Abstract


Objective: chloramphenicol as a highly effective broad-spectrum antibiotics, widely used for treatment of animals of various infectious diseases, strong because of its toxic side effects, many countries and regions have been prohibited or severely restricted the use of chloramphenicol, traditional chloramphenicol residue analysis method requires expensive equipment and professional and technical personnel, and long life cycle, high cost, and can not meet the requirements of the scene and quickly detect. Therefore, to establish a highly efficient, sensitive, economic chloramphenicol residue analysis method is necessary. Colloidal gold immunochromatographic strip assay specificity of antigen and antibody binding reaction measured trace antigen or antibody, the thesis aims at the development of the the chloramphenicol immune colloidal gold strip for used in livestock rapid detection of chloramphenicol residues in products, has important practical significance and the high value of commercial development. Methods: in vitro to expand the culture of hybridoma cells, the cells were inoculated into mice, the use of in vivo inducible prepared by a specific monoclonal antibody containing anti-chloramphenicol ascites. Ascites bitterness - ammonium sulfate, affinity column chromatography purification. Purified using antigen three monoclonal antibody from the protein concentration, potency, and cross-reactivity indicators the quality identification; using trisodium citrate reduction method different particle size colloidal gold, chloramphenicol specific immune The gold colloid uniform perfusion in the BT50-type glass fiber membranes, drying as soon as colloidal gold pad. Comparative Test selection the Whatman Company AE98Fast nitrocellulose membrane as a chromatographic membrane; control line using a 20-fold dilution of goat anti-mouse IgG; concentration 1mg/mL chloramphenicol package original (CAP-BSA) spraying detection line final assembly into the test strip. Results: select 4D10 antibody as a probe is prepared protein concentration 2.89mg/mL, measured by ELISA antibody titer to 1:1.024 × 106, the use of improved competitive ELISA determination of monoclonal antibody cross-reactivity with antibiotics, anti-chlorine ADM monoclonal antibody (CAP-McAb) cross-react with chloramphenicol sodium succinate no cross-reactivity with penicillin, streptomycin, sulfamethazine, thiamphenicol antibiotics with antibiotics, analogues The cross was less than 0.01%, with high specificity and affinity. Observed by UV analysis and the appearance of the test to determine the optimal protein concentration of colloidal gold-labeled antibody 48μg/mL, optimum pH about 8.2. Immunochromatographic strip obtained detect known sample containing chloramphenicol, sensitivity to 100ng/mL, reproducibility in its sensitivity range is 100%, the test strip clear bands appear in about 5min . Conclusion: The immune colloidal gold strip for the preparation of chloramphenicol validation tests show that the test strip detection time is short, good stability, in line with the requirements of the rapid detection of chloramphenicol specificity, simple operation , quick, preliminary scene bulk detection of chloramphenicol residues laid the foundation.

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CLC: > Medicine, health > Preventive Medicine,Health > Nutrition, hygiene,food hygiene > Food hygiene and food inspection > Food hygiene and inspection
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