ELISA - Definition and Overview

The Enzyme-Linked Immunosorbent Assay (ELISA or EIA for short) is a method usually employed in biochemistry to detect if a certain substance is present in a sample. It utilizes antibodies specific to the substance; these antibodies are linked to an enzyme which causes a chromogenic substrate to produce a signal.

The steps of the general ELISA test are:

1. Apply the sample to some sticky substrate, usually a plate with wells.
2. Apply the enzyme-linked antibodies and let them bind to the substance.
3. Wash the plate, so that unbound antibodies are removed.
4. Apply a chemical which is converted by the enzyme into a fluorescent signal.
5. View the result: if it fluoresces, then the sample contained the substance.

The enzyme acts as an amplifier: even if only few enzyme-linked antibodies are present, the enzyme molecules will produce many fluorescent signal molecules.

A variant of this technique is used in medicine to detect if a person's blood contains antibodies against a certain antigen (which would indicate past or present infection). The initial screening test for HIV infection is such an ELISA test. The steps are as follows:

1. Prepare a plate to which the antigen is bound
2. Apply the human serum to be tested
3. Wash the plate, so that unbound antibodies are removed.
4. Apply the enzyme-linked antibodies which specifically bind to the human antibodies.
5. Wash the plate, so that unbound enzyme-linked antibodies are removed.
6. Apply a chemical which is converted by the enzyme into a fluorescent signal.
7. View the result: if it fluoresces, then the serum sample contained antibodies against the antigen.

See also:

• Assay
• ELISPOT

External links

• An animated illustration of an ELISA assay (http://www.immunospot.com/elisa-animation.html)
• Online bookshelf from NCBI: searchable text books in molecular biology and related fields (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=Books)
• "Introduction to ELISA Activity - beginner walkthrough of ELISA used for detecting HIV, including animations (http://www.biology.arizona.edu/immunology/activities/elisa/main.html)
• Image of a plate, such as is commonly used in ELISA tests (http://www.freewebs.com/immunology/elisa-plate.html)