DEVELOPMENT OF A COMPETITIVE ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR DIETHYLCARBAMAZINE

A sensitive and reproducible competitive enzyme-linked immunosorbent a ssay (ELISA) for the determination of the concentration of diethylcarb amazine (DEC) in biological fluids was developed. Since DEC has no fun ctional group to conjugate with bovine serum albumin (BSA), N-(2-amino ethyl)-N-ethyl-4-methyl-1-pipe (DEC-NH2) was first synthesized. This c ompound was then converted to carboxyl DEC (DEC-COOH) and conjugated t o BSA and to poly-L-lysine for use as immunogen and sold-phase marker, respectively. The competitive ELISA was conducted by simultaneously i ncubating DEC with mouse anti-DEC antiserum over DEC-poly-L-lysine sol id phase. Subsequently, the binding of anti-DEC antibody was detected by using sheep anti-mouse IgG peroxidase conjugate as a tracer. The re liability. determined by the coefficient of variation for inter and in tra-assay, was satisfactory. The cross-reactivities of anti-DEC antibo dies with DEC metabolites, related compounds and ivermectin were negli gible. Using this assay, DEC levels were easily determined in serum of Mongolian jirds (Meriones unguiculatus) up to 4 hours following a sin gle dose of DEC citrate base(100 mg/kg of body weight) via intraperito neal route.