Development of an enzyme-linked immunosorbent assay for measurement of fire ant venom-specific IgE

An enzyme-linked immunosorbent assay (ELISA) was developed for in vitro measurement of IgE specific for Solenopsis invicta venom. Enhanced binding microtiter plates were coated with S. invicta venom protein and incubated with sera from fire ant allergic patients and control subjects. Bound IgE was tagged with peroxidase-conjugated monoclonal anti-IgE and quantitated with the substrate/indicator system H2O2/tetramethylbenzidine. Absorbance (620 nm) represented venom-specific IgE values. The ELISA correlated well with the imported fire ant venom RAST (r = .87, P < .0001). Using skin test reactivity as the standard measure of venom-specific IgE, the venom ELISA appeared to be a sensitive in vitro assay comparable to venom RAST. ELISA is less expensive than RAST and does not require licensing or handling of radioisotopes.