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Steps
Known antigen absorbed on 96 well plate
Patient serum added
If present: primary bodies bind to antigen
Enzyme linked secondary antibodies added → bind to Fc region of primary antibodies
Rinse plate & bound complex remain.
Substrate added → enzyme reacts → color change.
Look at results with Spectrophotometer
Testing for
HIV, HCV & HBV
Purpose
Detects unknown antibodies, IgG or IgM to diagnose viral infections.
Why sensitive
Multiple secondary ABs can bind to one primary AB which amplifies enxyme signal and improves detection even when antibody concentration is low.