Sensitive streptavidin-biotin enzyme-linked immunosorbent assay for rapid screening of chloramphenicol residues in swine muscle tissue.
A sensitive, competitive enzyme-linked immunosorbent assay (ELISA) for chloramphenicol (CAP) in swine muscle tissue has been developed. The ELISA is based on an earlier procedure. To improve sensitivity, different optimization procedures were investigated. The introduction of a streptavidin-biotin system and the use of a coating antigen with a lower CAP incorporation resulted in the most sensitive ELISA: the CAP concentration giving 50% inhibition decreased from 125 ng/mL to 3.0 ng/mL. This ELISA procedure was applied for a rapid screening of CAP residues in swine muscle tissue. The tissues were extracted with demineralized water. A concentrated phosphate-buffered saline solution was added to the filtered aqueous extract and this sample solution was directly submitted to the ELISA procedure. The results were compared to values obtained by analysis of a corresponding blank. This blank was prepared by treating a part of the aqueous sample solution with an immobilized monoclonal antibody preparation. This treatment was necessary because aqueous extracts of different swine muscle tissues showed a high variation in dose-response curves, probably caused by the complexity and variability of the matrix. In spiked tissues, the presence of CAP at concentrations of 10 micrograms/kg and higher can be easily demonstrated.
van de Water
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University of Utrecht, Faculty of Veterinary Medicine, Department of the Science of Food of Animal Origin, The Netherlands.
Haagsma
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D000469:Alkaline Phosphatase; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis*; D001710:Biotin / Q000032:analysis*; D002701:Chloramphenicol / Q000032:analysis*; D002846:Chromatography, Affinity; D004350:Drug Residues / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D007120:Immunochemistry; D007202:Indicators and Reagents; D009132:Muscles / Q000737:chemistry*; D019809:Streptavidin; D013552:Swine
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D007202:Indicators and Reagents; D002701:Chloramphenicol; D001710:Biotin; D019809:Streptavidin; D000469:Alkaline Phosphatase