Optimization of Elisa kit detecting chloramphenicol residues in aquaculture products

Abstract

Chloramphenicol is a wide-spectrum antibiotic. It proved to be specific activity against a wide variety of organisms that are causative agents of several diseases in domestic animals. Because of its serious adverse effects in humans, chloramphenicol has been banned within the European Union since 1994 for use in food-producing animals. According to the Vietnam Association of Seafood Exporters and Producers (VASEP), Japan has found residues of chloramphenicol in shrimp imported since February 2014. For this reason, the analytical method used to control chloramphenicol residues in livestock products is necessary. The application of the direct competitive enzyme-linked immunosorbent assay (ELISA) was optimized for the detection of chloramphenicol (CAP). Because the other instrumental methods included liquid chromatography-mass spectrometry (GC-MS), gas chromatography-mass spectrometry (GC-MS) appeared many disadvantages while enzyme immunoassays show some essential advantages such as quick, simple, economical, and sensitive. In this assay, the hapten chloramphenicol was constructed with a suitable enzyme to form enzyme conjugates by a modified active ester method. The anti-chloramphenicol antibodies were coated in microtiter plate for performance on an ELISA test kits. The aim of this study was to produce a suitable concentration of enzyme conjugates for the optimization of a sensitive and specific chloramphenicol ELISA. The IC50 values for chloramphenicol was 1 ppb and the limit of detection (LOD) was 0.06 ppb. The limit of quantification (LOQ) was 0.13 ppb and the recoveries ranged from 56.3% to 94.1%. Keywords: ELISA kit Chloramphenicol Enzyme conjugates