An enzyme-amplified diffraction-based immunoassay.

We have previously shown that a gold-conjugated secondary label can be used to reduce the limit of detection in a diffraction-based assay by more than 40-fold. We now show that by using a combination of a peroxidase-conjugated secondary label and a precipitating substrate the limit of detection in a diffraction-based assay can be reduced by more than 1000-fold. The response to secondary enhancement was linear for concentrations from 50 to 2000 pg/mL of antidigoxin.