CHAPTER -THREE
Immunohematology
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Immunohematology:
– Commonly known as "blood banking“
– Concept & techniques related to transfusion
therapy.
– Collecting, preparing, storing and transfusing blood.
– Immunologic reactions involving blood components
• Study of
– Red cell antigens and their corresponding antibodies
on blood
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Main application of immunohematology
Safe blood transfusion
Prevent of Rh immunization associated with
pregnancy
To prevent hemolytic transfusion reaction.
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Red Blood cell antigens
30 blood group systems with more than 200 unique red
cell Antigens.
• In biochemical terms red blood cell antigens may take the
form of:
Glycoprotein,
Glycolipids
proteins,
• Some of the red blood cell antigens are more
immunogenic
E.g. The D antigen within the Rh group system.
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Blood group Antibodies
Blood group antibodies blood group system are
classified into:
Natural and
Immune antibodies.
Natural / non red cell immune Abs
Are RBC Abs in the serum of an individual that are not
provoked by previous RBC sensitization.
• The term non-red cell stimulated is more
appropriate.
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– Because these Abs resulted from some
Non-red cell immune antibodies…
Characteristics
Are naturally occurring , produced from 37th day of fetal
life
They are mainly IgM type.
Exhibit optimum in vitro agglutination saline media: complete
antibodies.
Optimum reaction at room temperature or lower: cold
agglutinins.
Do not react above the body temperature
Most of these antibodies do not give rise to transfusion
reactions.
They are high molecular weight
6 Cannot cross the placenta
Immune antibodies
Produced due to previous antigenic stimulation
either by
transfusion or pregnancy
Characteristics
Mainly IgG type
Do not exhibit visible agglutination in saline,
but in albumin medium: Incomplete antibodies.
Optimally react at 370C: warm agglutinins.
Causes more serious transfusion reactions
than the naturally occurring ones.
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– Can cross the placental barrier.
Blood grouping
The ABO & Rh systems remain most important
transfusion therapy.
Blood typing is done
Before a person receives or donates blood
Before a person donates an organ for
transplantation
When a woman is planning to become pregnant
or first become pregnant
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ABO Grouping
ABO grouping has two components:
Forward(direct) grouping
Reverse(serum) grouping
Principle
When an antigen is mixed with its
corresponding antibody under the right
conditions it causes agglutination or
hemolysis of the red cells
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forward grouping or Direct (cell grouping)
Testing of the red blood cells for the presence of ABO
Antigens .
RBCs + Antiserum Agglutination
(Ag) (Rgt) ( Ag-Ab rxn )
Interpretation
+ve agglutination with specific antiserum indicates the
presence of Ag
-ve agglutination indicates the corresponding Ag is not
present .
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Reverse grouping or Indirect (serum
grouping)
Testing of serum or plasma for the expected ABO
Antibodies (or).
Serum + RBCs +ve agglutination
Interpretation
Positive agglutination:- Abs specific for Ag are
present.
Negative agglutination:- No Abs are present for Ags.
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Clinical significance of blood grouping
For safe blood transfusion
Prevention of Hemolytic Disease of the
Fetus and Newborn (HDFN)
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Points to be considered in routine
phenotyping
Donor and recipient red blood cells must be tested
using anti- A and anti-B.
Donor and recipient serum or plasma must be tested
for the expected ABO antibodies using reagent A 1 and
B red blood cells.
Cord blood and samples from infants less than 4
months old should be tested by forward ABO grouping
.
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The ABO phenotype is determined when the
red blood cells are directly tested for the
presence or absence of either A or B antigen.
Serum testing provides a control for red
blood cells testing, since ABO antibodies
would reflect Landsteiner’s rule.
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phenoty RBC reaction or serum or plasma
pe direct blood reaction reverse
grouping blood groupng
Anti-A Anti-B A cell B- cell
A + 0 0 +
B 0 + + 0
AB + + 0 0
O 0 0 + +
+=agglutination 0= no agglutination
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Rh- Blood Group System
There are over 40 known Rh Ags, five of w/c
commonly cause post transfusion problems
(D,C, E, c, e).
The antigen is well developed before birth.
Rh-Antibodies
No naturally occurring anti-D Abs
IgG in type, can pass through placenta.
They are clinically significant because the Rh( D)
17 antigen is potent immunogen
Rh- typing
The importance of Rh typing
Administration of Rh +ve blood to Rh –ve may
sensitize the person to form anti-D antibody
Donation of Rh +ve blood to recipient having anti-D
could be fatal .
RBCs + Anti-D + agglutination
This can be performed on slide test , saline tube test.
Interpretation
Agglutination of RBCs --positive for D-Ag( Rh
+ve )
No agglutination of RBCs---the D-Ag is not expressed (Rh-
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ve)
THANK YOU !
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