Detection of Ribavirin Residues in Animal-Derived Food by Way of Direct Competitive ELISA

Aiming at the problem of ribavirin residues in animal source foods,a direct competitive enzyme-linked immunosorbent assay(dc-ELISA)was established to detect ribavirin residues rapidly and efficiently. Ribavirin(RBV)was conjugated with keyhole limpet hemoeyanin(KLH)by active ester method to prepare the ribavirin complete antigen(KLH-RBV). The antiserum was purified with Protein A-Sepharose 4B to prepare polyclonal antibody against ribavirin. A dc-ELISA was developed to detect and quantitate ribavirin with IC₅₀ of 10.84 ng/mL and detection limit of 0.002 ng/mL. This method was applied to determine RBV in real chicken,chicken liver,pork and pig liver samples with satisfactory result of recovery rate ranged from 80.23% to 90.07%. HPLC was used to verify the results and the results of the two methods showed a good linear relationship. The method established was sensitive and the procedure of sample pretreatment was simple and quick. It was suitable for spot rapid detection of RBV residue in animal source foods.