MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

GROUP MEMBERS: KHALIPHA N 201614708

MHLANGA S. 2018

MBOZI S 2019

NGQAKAYI A. 2019

DYANI L.Q 2017

COURSE CODE: MIC 312

TITLE: AGGLUTINATION

TUTOR’S NAME:

DUE DATE: 26/05/2021

1
MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

Contents
AGGLUTINATION............................................................................................................................3
TYPES OF AGGLUTINATION REACTIONS................................................................................3
ACTIVE AGGLUTINATION............................................................................................................3
Slide/ tile agglutination....................................................................................................................3
Tube agglutination...........................................................................................................................3
Heterophile agglutination test.........................................................................................................3
Antiglobulin (Coombs) test.............................................................................................................4
PASSIVE AGGLUTINATION...........................................................................................................4
Latex agglutination..........................................................................................................................4
HEMAGGLUTINATION TEST........................................................................................................5
Viral hemagglutination...................................................................................................................5
Coagglutination test:.......................................................................................................................5
Uses of Coagglutination test........................................................................................................5
REFERENCES....................................................................................................................................8

2
MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

Agglutination is an antigen-antibody reaction whereby a particulate antigen reacts with

an antibody leading to visible clumps known as agglutination. Briefly discuss the types
of agglutination reactions and give examples for each.

AGGLUTINATION
According to Cole (et al, 2019), Agglutination is the visible expression of the aggregation of
antigens and antibodies. Agglutination reactions apply to particulate test antigens that have
been conjugated to a carrier. However, if an antigen is present in a test specimen, it will bind
to the antibody and form visible, cross-linked aggregates.

TYPES OF AGGLUTINATION REACTIONS

ACTIVE AGGLUTINATION
Agglutination reaction where the antigens are discovered normally on a molecule are known
as direct agglutination. In active agglutination, direct agglutination of particulate antigen with
specific antibody occurs. Direct bacterial agglutination utilizes entire microbes as a source of
antigen. It measures the antibody level delivered by a host infected with that microbe (Cole et
al, 2019). The binding of antibodies to surface antigens on the bacteria brings about visible
clumps, Active agglutination can be of following types:

Slide/ tile agglutination

basic types of agglutination reaction that is performed on a slide. identification of bacterial
types represents an exemplary illustration of a slide agglutination. In this strategy suspension
of unknown antigen is kept on slide and a drop of standardized antiserum is added or the
other way around. A positive reaction is demonstrated by development of noticeable clumps.
For example, Widal test, RPR test (Cole et al, 2019).

3
MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

Tube agglutination
It is agglutination test performed in tube and standard quantitative technique for
determination of antibody titre. In this method serum is diluted in a series of tubes and
standard antigen suspensions (specific for the suspected disease) are added to it. After
incubation, antigen-antibody reaction is indicated visible clumps of agglutination (Roelcke,
2016).

Heterophile agglutination test

This test depends on demonstration of heterophilic antibodies in serum present in certain

bacterial infections.

Antiglobulin (Coombs) test

This' test was devised by Coombs, Mourant, and Race for identification of fragmented anti-
Rh antibodies that do not agglutinate Rh+ erythrocytes in saline. At the point when serum
containing incomplete anti-Rh antibodies is mixed with Rh+ erythrocytes in saline,
incomplete antibody antiglobulin coats the outside of erythrocytes yet does not bring about
any agglutination. At the point when such erythrocytes are treated with antiglobulin or
Coombs’s serum (rabbit antiserum against human gamma globulin), at that point the cells are
agglutinated. Coombs’s test can be direct as well as indirect (Roelcke, 2016).

In direct method, the sensitization of red blood cells (RBCs) with incomplete antibodies takes
place in vivo. Cell-bound antibodies can be detected by this test in which antiserum against
human immunoglobulin is used to agglutinate patient’s RBC. In indirect method, the
sensitization of RBCs with incomplete antibodies takes place in vitro. Patient’s serum is
mixed with normal red cells and antiserum to human immunoglobulin. Agglutination occurs
if antibodies are present in serum. Coombs’s test is used for detection of anti-Rh antibodies
and incomplete antibodies in brucellosis and other diseases (Mailafia et al, 2019).

PASSIVE AGGLUTINATION
Passive agglutination employs carrier particles that are coated with soluble antigens. In this
either antibody or antigen is attached to certain inert carrier thereby, particles or cells gets
agglutinated when corresponding antigen or antibody reacts. Latex particles, Carbon
particles, Bantonite etc. are used as inert carriers. E.g., Antigens coated in latex particles used

4
MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

in ASO test. When the antibody instead of antigens is adsorbed on the carrier particle for
detection of antigens, it is called reverse passive agglutination (Cole et al, 2019).

Latex agglutination
It employs latex particles as carrier of antigen or antibodies. In latex agglutination, many
antibody or antigen molecules are bound to latex beads (particles), which increases the
number of antigen-binding sites. If corresponding antigen or antibody is present in a test
specimen, antigen antibody binds and form visible, cross-linked aggregates. Latex
agglutination can also be performed with the antigen conjugated to the beads for testing the
presence of antibodies in a serum specimen (Maltezou et al, 2020).

HEMAGGLUTINATION TEST
RBCs are used as carrier particles in hemagglutination tests. RBCs of sheep, human, chick,
etc. are commonly used in the test. When RBCs are coated with antigen to detect antibodies
in the serum, the test is called indirect hemagglutination (IHA) test. Hemagglutination uses
erythrocytes as the biological carriers of bacterial antigens, and purified polysaccharides or
proteins for determining the presence of corresponding antibodies in a specimen. When
antibodies are attached to the RBCs to detect microbial antigen, it is known as reverse passive
hemagglutination (RPHA) (Mailafia et al, 2019).

Viral hemagglutination
Many viruses including influenza, mumps, and measles can agglutinate RBCs without
antigen–antibody reactions. This process is called viral hemagglutination. This
hemagglutination can be inhibited by antibody specifically directed against the virus, and this
phenomenon is called hemagglutination inhibition (Roelcke, 2016).

Coagglutination test:
Coagglutination is a type of agglutination reaction in which Cowan I strain of S. aureus is
used as carrier particle to coat antibodies. Cowan I strain of S. aureus contains protein A, an
anti-antibody, that combines with the Fc portion of immunoglobulin, IgG, leaving the Fab
region free to react with the antigen present in the specimens. In a positive test, protein A
bearing S. aureus coated with antibodies will be agglutinated if mixed with specific antigen.
The advantage of the test is that these particles show greater stability than latex particles and
are more refractory to changes in ionic strength (Cole et al, 2019).

5
MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

Uses of Coagglutination test

 Detection of cryptococcal antigen in the CSF for diagnosis of cryptococcal
meningitis.
 Detection of amoebic and hydatid antigens in the serum for diagnosis of amoebiasis
and cystic echinococcosis,
 Grouping of streptococci and mycobacteria and for typing of Neisseria gonorrhoeae.

B)Latex agglutination is one of the agglutination reactions whereby a specific antigen is

mixed with an antibody that is coated on the surface of latex particles. Briefly discuss
the use of this reaction in detection of E. coli 0157:H7 pathogen.

E. coli 0157:H7 is a pathogen that affects both animals and humans. When a latex
agglutination test is performed to test for E. coli 0.157:H7, two drops of saline are placed on a
reaction card using a micropipette. Some of the E. coli culture is collected and mixed
thoroughly with the two drops of saline on the reaction card using a mixing stick. Then one
drop of 0157 test latex is placed in each of the drops of saline that contain the E. coli culture,
where one of the drops serves as the control, and mixed thoroughly using a mixing stick. The
reaction card is then rocked carefully for 30 seconds and then it is observed for agglutination.
The agglutination shows that there is a presence of 0157 antigens in the test culture (Mailafia
et al., 2017).

Test cultures that are positive for 0157 antigen are then further tested for H7 antigen. On a
reaction card, two circles of the culture are placed and H7 test latex was placed on each circle
of culture. The contents of the culture were then mixed thoroughly using a mixing stick. The
reaction card was then rocked carefully for 30 seconds and it was observed for agglutination.
The presence of agglutination of the contents of the reaction card shows the presence of the
H7 antigen (Mailafia Et al., 2017).

6
MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

Figure 1: Reaction card showing the agglutination of the 0157 test latex, agglutination of the
H7 test latex and the control.

C)Immunisation is one of the key methods used to control viral infections. Discuss
immunisation regarding COVID19.

Immunisation is the process by which an individual's immune system becomes fortified

against an agent. Vaccines train your immune system to create antibodies, just as it does
when it is exposed to a disease. When you get a vaccine, your immune system responds. It
recognises the invading germ, such as the virus or bacteria produces antibodies. Antibodies
are proteins produced naturally by the immune system to fight disease remembers the disease
and how to fight it. If you are then exposed to the germ in the future, your immune system
can quickly destroy it before you become unwell. However, because vaccines contain only
killed or weakened forms of germs like viruses or bacteria, they do not cause the disease or
put you at risk of its complications (Roelcke, 2016).

In the early months of 2020 when the World Health Organisation first declared covid-19 a
pandemic, it was encouraged that people get the influenza vaccine. It was shown by

7
MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

mathematical models that by being vaccinated with the influenza vaccine, it would help to
manage the symptoms of covid-19 during the influenza season (Maltezou et al., 2020).

The vaccination uses your body’s immune system to increase protection to an infection
before you meet that infection. In other words, it is like being infected with the disease
without suffering the actual symptoms. If you meet an infection after you’ve been vaccinated,
your body works to stop you from getting the disease, or you may get just a mild case. Unlike
other proposed approaches to immunisation (such as homeopathy), vaccinations have been
rigorously tested to demonstrate their safety and effectiveness in protecting against infectious
disease (Maltezou et al, 2020).

REFERENCES

 Cole, J.R., Sulzer, C.R. and Pursell, A.R., (2019). Improved microtechnique for the
leptospiral microscopic agglutination test. Applied microbiology, 25(6), pp.976-980.

 Mailafia, S., Madubuike, S. A., Raji, M. A., Suleiman, M. M., Olabode, H. O. K.,
Echioda-Egbole, M. and Okoh, G. P. R., (2017), Phenotypic identification of
Escherichia coli 0157:H7 isolates from cattle at Suleja Abattoir, Nigeria, African
Journal of Microbiology Research, 11(21), pp.845-850.

 Maltezou, H. C., Theodoridou, K. and Poland, G., (2020), Influenza immunization

and covid-19, American Society of Internal Medicine, 38(39), pp. 6078-6079.

8
MIC 312 ASSIGNMENT 3 GROUP 1 26/05/2021

 Roelcke, D., (2016). Cold agglutination. Transfusion medicine reviews, 3(2), pp.140-
166.