Demonstration of ELISA Test

An enzyme-linked immunosorbent assay, also called ELISA or EIA, is a test that detects
and measures antibodies in your blood. This test can be used to determine if you have
antibodies related to certain infectious conditions. An ELISA test uses components of the
immune system (such as IgG or IgM antibodies) and chemicals for the detection of immune
responses in the body (for example, to infectious microbes). The ELISA test involves an
enzyme (a protein that catalyzes a biochemical reaction). It also involves an antibody or
antigen (immunologic molecules). Examples of the uses of an ELISA test includes to
diagnose infections such as HIV (human immunodeficiency virus) and
some allergic diseases like food allergies. ELISA tests are also known as an immunosorbent
assay.
What is the use of an ELISA test?

ELISA tests are primarily used for the detection of proteins (as opposed to small molecules
and ions such as glucose and potassium). The substances detected by ELISA tests can
include hormones, viral antigens (dengue fever, for example), bacterial antigens (TB, for
example), and antibodies that the body has made in response to infection (antibodies
to hepatitis B, for example) or vaccination.

What is an ELISA kit?

An ELISA kit is a commercially available ELISA test that usually contains pre-coated
polystyrene plates, detection antibodies, and usually all of the chemicals needed to perform
an ELISA test. However, special kits can be purchased with substances designated by the
customer.

How do health care workers perform an ELISA test?

Health care personnel who perform the test are trained laboratory technicians who use
special kits that measure the antigens' interactions with the antibodies in the kit. They will
inform your doctor of the test results.

How does ELISA testing work?

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There are variations of the ELISA test (see below), but the most utilized type consists of an
antibody attached to a solid surface (polystyrene plate). This antibody has affinity for (will
latch on to) the substance of interest, such as a hormone, bacteria, or another antibody. For
example, human chorionic gonadotropin hormone (HCG), the commonly measured protein
which indicates pregnancy, can be detected by ELISA. A mixture of purified HCG linked to
an enzyme and the test sample (blood or urine) are added to the test system. If no HCG is
present in the test sample, then only the linked enzyme will bind to the solid surface. The
more substance of interest that is present in the test sample, the less linked enzyme will
bind to the solid surface. The more of the substance of interest is present it will cause a
reaction and show up on the test plate in some way, such as a change in color of the
solution (or like a pregnancy test "two pink lines" or a "+" mark).

What are the types of ELISA tests? What is a direct ELISA?

There are four types or kinds of ELISA tests:

• Direct ELISA: attachment of an antigen to a polystyrene plate followed by an

enzyme-labeled antibody that can react with the antigen and a substrate that can be
measured
• Indirect ELISA: attachment of an antigen to a polystyrene plate followed by an
unlabeled or primary antibody followed by an enzyme-labeled antibody that can
react with both the primary antibody and substrate
• Sandwich ELISA: A capture antibody is attached to the polystyrene plate, then
antigen is added that specifically attaches or captures the antigen. A second
antibody, also specific for the antigen but not the same as the capture antibody is
added and "sandwiches" the antigen. This second antibody is then followed by an
enzyme-labeled antibody specific for the second antibody that can react with a
substrate that can be measured
• Competitive ELISA: This test is like the sandwich ELISA but involves the addition of
competing antibodies or proteins when the second antibody is added. This results in
a decrease in the substrate signal that is generated. This test is considered to give
good, highly specific results.

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What are the advantages of ELISA testing?

ELISA tests are generally good and accurate tests. They are considered highly sensitive and
specific (accurate) and compare favorably with other methods used for the detection of
substances in the body. The ELISA testing method is more straightforward and easier to
perform than older laboratory techniques, which often required radioactive materials.

n general, people do not need to prepare for an ELISA test. The test is done in a lab. If your
blood is required, the only hurt is in blood collection. The risks associated with an ELISA
test are rare and associated with blood withdrawal (infection, vessel damage, for example).

Depending on what the test is being used for, you may get results as quickly as about 24
hours if the test is done locally. However, there are some tests that may take days to weeks.

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