NAME: MECHELLE SIBANDA

STUDENT NUMBER: N02219016F

COURSE: INTRODUCTION TO IMMUNOLOGY
CLASS: SBB
LECTURER: dr ZUMBIKA
COURSE CODE:SBB2104

TITLE: IMMUNOCHROMATOGRAPHIC ASSAY

INTRODUCTION
Immunochromatography assay is an analytical device used to used to detect the presence of
one or more target analytes in a liquid sample. The device is used to test for many substances.
It can test for pregnancy , Human Immunovirus HIV andinfectious diseases such as malaria,
dengue fever, and tuberculosis. It is a simple, easy-to-use, and cost-effective test method that
does not require sophisticated laboratory facilities or equipment. The test is designed to
provide accurate and reliable results within minutes, making it ideal for point-of-care testing
and outreach programs that focus on under served populations. This method is a combination
of immunochemical reactions and chromatography. Chromatography is a physical method of
separation of the components of a mixture by a continuous redistribution between two
phases, one stationary and the other moving. A variety of attractive forces between the
stationary phase, and the substance to be separated lead to the selective retardation of the
latter, relative to the moving phase.
A typical immunochromatographic test strip consists of an absorbent pad onto which
specimen is applied, a conjugate or reagent pad that contains antibodies specific to the target
analyte conjugated to colored particles (i.e., colloidal gold particles or latex microspheres), a
reaction membrane onto which anti target analyte antibodies are immobilized in a line across
the membrane as a capture zone or test line, a control zone containing antibodies specific for
the conjugate antibodies, and a waste reservoir composed of another absorbent pad designed
to draw the sample across the reaction membrane by capillary action.

Image 1
The above image shows lateral flow to detect an antigen. The specimen containing the
antigen (Analyte) is placed on the sample pad. The antigen, with the fluid, moves to the
conjugate pad where it is bound by a labelled antibody specific to the targeted antigen. The
conjugate pad also contains labelled antibodies nonspecific to the antigen to be detected.
Antigen-antibody complexes migrate through the nitrocellulose membrane and reach the
'Test line' area. In this area, antigen-specific antibodies are immobilized to catch the antigen-
antibody complexes. When antigen-antibody complexes accumulate in this area, the line
becomes visible to the naked eye. The non-specific antibodies also migrate and pass the 'Test
line' to reach the 'Control line' area and that line also becomes visible. The test is considered
positive only when the two lines (T and C) are visible.
This is how most pregnancy test are carried out. This test is an antigen-capture
immunochromatographic assay. In that test the hormone human chorionic gonadotropin
(hCG)

Rapid antigen tests are quicker, but they require a higher viral load to produce accurate
results.Immunochromatography HIV tests have some limitations and drawbacks. The test
requires a detectable level of HIV antibodies, meaning it may produce a false negative result
during the acute phase of the infection. The test may also produce an indeterminate result,
requiring further testing to confirm the diagnosis.
Another limitation of immunochromatography HIV tests is that they cannot distinguish
between HIV-1 and HIV-2. This is important because HIV-2 is less common and has
different treatment options. Additionally, the test may produce false positive results due to
cross-reactivity with other antibodies, such as those from a recent flu vaccine or autoimmune
disorders. It is important to confirm positive results with additional testing to avoid
unnecessary anxiety and treatment.

Laboratory diagnosis of SARS-CoV-2: available approaches and limitations - Scientific

Figure on ResearchGate. Available from: https://www.researchgate.net/figure/Principle-of-
immunochromatography-This-drawing-shows-lateral-flow-to-detect-an-
antigen_fig2_342166317 [accessed 6 Nov, 2023]

Performance Characteristics of a Rapid New Immunochromatographic Test for Detection of

Antibodies to Human Immunodeficiency Virus - Scientific Figure on ResearchGate.
Available from: https://www.researchgate.net/figure/Immunochromatographic-tests-Rapid-
Check-HIV-1-2-and-Determine-used-for-the-detection-of_fig1_8041160 [accessed 6 Nov,
2023]