Example of an indirect ELISA test: testing for antibodies
An indirect ELISA test can be used to test whether a patient has?antibodies?to?HIV:
First, HIV?antigens?are?bound?to the bottom of the reaction vessel
A blood plasma sample is then taken from the patient and added to the reaction vessel
Any?HIV-specific antibodies?(i.e. the antibodies produced against HIV) that are present in the blood plasma now?bind?to the HIV antigens (stuck to the bottom of the reaction vessel). These HIV-specific antibodies are known as the?primary antibodies
Any other antibodies that are present in the blood plasma are?unbound?and are?washed out
A second type of antibody with an?enzyme attached?to it is added to the reaction vessel. These are known as the?secondary antibodies
These secondary antibodies?bind?to the primary antibodies. The reaction vessel is?washed out again?to?remove?any?unbound?secondary antibodies. This is a very important step in?avoiding false-positive test results. If they are not washed out, unbound secondary antibodies would give a positive result, even if there were?no?primary (HIV-specific) antibodies present to start with
Finally, a solution is added that contains a?substrate?that reacts with the enzyme attached to the secondary antibodies. If there are any secondary antibodies present, a?coloured product?is formed, causing the solution in the reaction vessel to?change colour. This indicates that the patient has HIV-specific antibodies in their blood (and therefore they are infected with HIV)
An indirect ELISA test can be used to test whether a patient has antibodies to HIV
Example of an indirect ELISA test: testing for antigens
Another type of indirect ELISA test can be used to test whether a patient has specific?antigens?in their blood plasma
For example, this can be used to test for?prostate cancer:
Prostate cancer is a cancer of the prostate gland (so only occurs in men)
The blood plasma of a patient can be tested for the presence of?prostate-specific antigens?(PSAs)
If the PSA concentration of the blood plasma is abnormally?high, this suggests that the patient has prostate cancer and further diagnostic tests will be carried out
This indirect ELISA test is very similar to the one described for HIV above
The main difference is that?antibodies to PSA?are bound to the bottom of the reaction vessel (instead of antigens, as in the HIV example)
An indirect ELISA test can be used to test whether a patient has prostate-specific antigens