Practical Assessment on Blood Grouping Techniques

The presence of antigens and antibodies in the blood plasma plays a fundamental aspect when conducting blood transfusion matching. A mixture of blood type groups causes serious problems which lead to agglutination and eventual death due to lack of compatibility between blood components. The Rhesus factor is a fundamental aspect which plays a key role in the production of antigens when the blood group is not compatible.

Running head; Practical Assessment on Blood Grouping Techniques
UNIVERSITY
Task
The investigation into blood grouping techniques and implications in clinical Immunology
and Immunohaematology
Student Name
Tutor

Clinical Immunology and Immuno haematology 2
Abstract
The presence of antigens and antibodies in the blood plasma plays a fundamental aspect when
conducting blood transfusion matching. A mixture of blood type groups causes serious
problems which lead to agglutination and eventual death due to lack of compatibility between
blood components. The Rhesus factor is a fundamental aspect which plays a key role in the
production of antigens when the blood group is not compatible. Performing blood tests is key
in establishing the compatibility process. ABO forward, reverse and D typing are a key
process performed to assess compatibility tests of blood before transfusions take place. This
practical experiment performed centrifuge separation for the different blood groups. The
practical utilized aspects of forwarding, reverse and RhD typing to blood samples. The results
obtained showed how different phenotype produces different genotypes among a family of 6
individuals entailing, parents both male and female and four daughters. The results yielded
various genotypes as observed from the daughter's blood groups. Further, the Rhesus factor
was established and found 3 blood specimen having Rh D negative antibodies. Blood
grouping analysis is thus crucial in determining the compatibility of individuals in receiving
different blood group types.
Introduction
The scientific understanding of the blood non-universality principle is due to the immune of
our systems as they produced antibodies against each type (Hayden et al., 2006). The four
groups of blood are identified genetically. The difference occurs due to protein situated on
the surface of red blood cells which act on antigens, also referred to as agglutinogens and
plasma proteins which acts as antibodies referred to as agglutinins. Blood type A has A
antigens situated on the surface of the red blood cells and B antibodies situated in the blood
plasma. Blood type B contains B proteins situated on the surface of the red blood cells and
anti A antibodies in the plasma (Malomgre & Neumeister, 2009).
The three major tests often performed on the individual to ascertain individual ABO/D type
include ABO forward, ABO reverse and D typing. ABO typing refers to the determination of
the presence of antigen A and B antigens in blood plasma. Reagent anti-A and Anti B sera is
essential in this aspect. Agglutination signifies antigen presence on a red cell while lack of
agglutination signifies its absence (Flower, Roulis & Hyland, 2018).
ABO reverse typing is used to initiate detection of ABO antibodies on an individual serum. It
is used to ascertain the ABO forward typing. The serum of the patient is mixed with reagent
group A cells (Huet, Cubizolles & Buhot, 2018). Serum outcome serum typing or reverse

Clinical Immunology and Immuno haematology 3
typing is often compared with forwarding typing to facilitate accurate ABO determination
(Then et al., 2015).
Centrifuge application has been applied in with the role of centrifuge in blood testing being to
separate blood components. For each component, blood typing was centrifuged in a tube,
(Gani et al., 2015 pp 70). At this stage, blood can be separated into its various components
which include red blood cells, white blood cells, and plasma.
The aim of this practical experiment is geared towards introducing blood grouping techniques
into practice. Further performance of centrifuge will be performed on the blood. A further
key aspect of determination is the principles of forward and reverse ABO grouping and RhD
typing which will be used to facilitate blood interpretation.
Materials and Methods
1 bottle containing 10 ml of antigen A, B, A, Band D, universal 3% suspension of blood
group A and B red cells, pasteur pipette, test tubes and marker pen and rack were used this
assessment. Setting of ABO cell and serum typing and RhD type for parent 1, daughter D1
and Daughter D2 labelling was done. The tubes were labelled with accurate signage having
anti-A, anti-B, anti-A,B and anti-D. The appropriate test cell suspension was added and
incubated in the bench for 5-10 minutes the tubes spanned at 1000rpm for 20 seconds.
In serum grouping, A and B cells were labeled. 2 drops of parent plasma were added with 1
drop of reagent test cells –A and B into the labeled tubes. The cells and the serums were
mixed and incubated for 8 minutes and centrifuged tubes at 1000rpm for an estimate of 20
seconds at room temperature.
Results
The results obtained from the experiment were recorded and tabulated in the table below and
their interpretation on serum group interpretation and full blood type interpretation as
observed in table 1 below.
Parent
/offspring
Number
Id Number Antisera Cell Group
Interpretation
Reagent
Cells
Serum Group
Interpretation
Antiser
a
Full
Blood
type
Interpreta
tion
XXXX X XXXXXXX -
A
-
B
-A,
B
XXXX A B XXXX -D (ABO and
RhD)
Parent 1 4 4 4 AB 0 0 -Negative 3 AB(O) D+
Parent 2 4 0 4 A 0 3 Beta 0 A() D-
Daughter 1 4 0 4 A 0 4 beta 0 A() D-
Daughter 2 4 4 3 AB 0 0 negative 4 AB(O) D+

Clinical Immunology and Immuno haematology 4
Daughter 3 4 0 4 A 0 4 beta 0 A() D-
Daughter 4 0 0 0 O 3 4 Alpha&beta 0 O (&)
D-
Discussion
Blood types are grouped in several ways. The basic system is referred to as ABO, which
offers a division of four blood groups, that are distinct groups; A, B, AB, and O. Every
individual blood contains an inherited set of antigens. Anti D antigen control is essential in
AB positive patient. If a patient is AB positive, forward typing tube will not take place with a
negative reaction. Negative control tube indicates the individual state of AB positive while
positive reaction indicates the need for more testing to determine the ABO and D type of the
person, (Selleng et al., 2017). From the results, it indicates that parent 1 has blood group AB
while parent 2 and daughter 1 have same blood group while daughter 2 has AB, daughter 3
has blood group A while daughter 4 is a universal donor.
Rh factor is a significant aspect of blood grouping. The key significant aspect is the D antigen
which is a factor on the immune system. D negative don has ant D IgG antibodies. D-
negative individuals can produce IgG anti D antibodies (Fasano, Ross & Stella, 2016).
Patient with AB positive indicates no reaction in the tube during forwarding type with a
negative reaction. Even if the reverse type reaction matches, no agglutination takes place
between A and B red blood cell. Positives which might occur in forwarding tubes could be
attributed to spontaneous agglutination of red blood cells as a result of protein coating of red
blood cells (Gildersleeve & Wright, 2016 pp 445).
D negative result test is geared towards controlling the AHG phase of testing and prevent
detection failure of positive reaction due to patient red blood cell being coated with antibody.
There is complete and strong agglutination depicted from antisera for the patient red blood
cells as indicated in the table above giving rise to corresponding blood group typing
(Tchakarov, Hobbs & Bai, 2014).
Conclusion
ABO blood grouping systems entail two antigen and antibodies found in the human blood.
Antigen A and B are the two common antigens in the blood. Present antigens are located on
the red blood cells and serum of antibodies. This investigation has analyzed blood group for
various patients and results has shown the various classifications and their corresponding
antigens (Barty et al., 2017). Agglutination reaction assessment is key before any transfusion

Clinical Immunology and Immuno haematology 5
process and it occurs between similar antigen and antibodies. Rh system is a critical factor on
blood group system. The key significant factor is the D antigen. The D negative persons do
not have any antibody production; however, they can produce anti D antibodies (Quraishy &
Sapatnekar, 2016 pp 220).
Interpretive exercise
The possible genotype of patient 1
Phenotype AB
Possible genotype is AB
The possible genotype of patient 2
Phenotype A
The possible Genotype is AA or AI
Family tree inheritance of ABO antigens
AB A
A AB A O
Impact of Rh-positive blood transfusion on blood group type A
Blood cells have combination of important substances referred to as antigens found on their
surfaces. Antigens perform important function on the immune system which enable it to
distinguish it from own body cells and other foreign invaders. When foreign matter is
established on the system, it is destroyed by the antibodies. Antibodies attach themselves on
the red blood cells which bind them. Agglutination occurs when the red blood cells are
bounded together, thus it is critical for donors red blood cells to be checked so as to alleviate
agglutination during the transfusion.
There is
strong
agglutination
occurring on
antibody A
There is strong
agglutination
occurring
between A and
B
There is
strong
agglutination
occurring on
antibody A
There is no
agglutination
occurring
hence universal
blood group

Clinical Immunology and Immuno haematology 6
Rehsues factor is a critical aspect during blood transfusion process. Patients or individuals
having Rh positive have D antigens on its surface of the red blood cells while the Rh negative
individuals do not have them. The ABO group, antibodies often repel against D antigens
which are present in normal conditions, (Yeow et al., 2016). When an Rh-negative receives a
transfusion from Rh+, it leads to the production of antibodies by producing ant D which leads
to agglutination (Moschini et al., 2016).
When transfused with blood group type A, the individual develop anti B antibody plasma,
this could lead to agglutination of red blood cells of the person. The patient will be
compatible with donor blood group O and A while they will incompatibility with B and AB
(Franchini, Mengoli & Lippi, 2016). The occurring sensitization of the Rh D antigens could
lead to potential production of maternal IgG antibodies which can be passed through the
placenta. Hence the development of Rh antibodies is likely to lead to the production of IgG
antibodies from the exposure occasioned by Rh-positive transfusion (Meyer & Uhl, 2015.
The implication if a woman has Rh- and is a pregnant with a Rh+ foetus, is that the first
pregnancy has no harm as the mother might be exposed, as the fusion of the Rh+ and Rh-
blood during either birth or miscarriage, cannot pose any danger. Antibodies are not produced
during the first exposure, however during the second conception; anti D antibodies can pass
through the placenta and form agglutination which endangers the foetus growth, thus it will
be essential to provide immune globulin during 28-32 weeks of gestation and at birth.

Clinical Immunology and Immuno haematology 7
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