List the steps in an indirect solid phase immunosorbent assay for Hepatitis B surface
antigen (HBsAg) using an enzyme labeled detection antibody.
What will be an ideal response
1. Obtain solid phase (tube, microtiter well, bead, etc.) coated with anti-HBsAg.
2. Add specimen to solid phase. Incubate. Wash off excess specimen.
3. Add anti-HBsAg from an animal (goat, mouse, etc.). Incubate. Wash off excess.
4. Add enzyme-labeled animal antibody (e.g., anti-goat IgG). Incubate. Wash off excess enzyme-labeled animal
antibody.
5. Add enzyme substrate and chromogen. Incubate until sufficient color is formed and quantify color.
6. Highly colored specimens are positive for HBsAg.
A direct solid phase immunosorbent assay would be identical except step 3 would be to add anti-HBsAg labeled
with an enzyme and step 4 would not be done.
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