Abstract
Phenobarbital was detected by an enzyme-linked immunosorbent assay (ELISA) using an avidin-biotin-peroxidase complex (ABC) technique. The antigen, p-succinamidopheno-barbital conjugated to bovine serum albumin, was adsorbed to wells of a microtiter plate made of polystylene. Rabbit anti-phenobarbital antiserum, samples, biotinylated anti-rabbit IgG antibody, and an ABC reagent were then added to the wells. The absorbance produced by the interaction of the peroxidase activity with a chromogenic substrate was measured at 492 nm.
Less than 1 ng of phenobarbital could be detected by this procedure, and the color development was linearly inhibited up to the addition of 20 ng of phenobarbital. Good correlation between this ELISA and the corresponding radioimmunoassay (RIA) was obtained (r=0.95).
