ABO and Rh Typing

Composition of Blood
ANTIGEN – substances that induces an immune
response by causing production of antibodies

ANTIBODY - serum protein that is induced by, and

reacts specifically with a foreign substance (Ag)
 ABO Blood Group System
• Discovered by Karl Landsteiner; on chromosome 9

• Single most important blood group for the selection and

transfusion of blood

• Three antigens: A, B, H

• Two major antibodies: anti-A and anti-B

• Four phenotypes: A, B, AB, O

 ABO Antigens
Present on the surface of red cells as well as tissue and
endothelial cells in the body

Found in soluble form in the plasma and other body

secretions in people known as secretors

3 possible genes tat an be inherited: A, B, and O

 ABO Antibodies
Natural antibodies antigenic stimulus is
environment exposure occur at birth

Newborns without ABO antibodies of their own;

begin to produce wit detectable titers at 6 months age
ABO blood grouping system

According to the ABO blood

typing system there are four
different kinds of blood types:
A, B, AB or O (null).

 
AB0 blood grouping system

Blood group A
If you belong to the blood
               group A, you have A antigens
on the surface of your RBCs
and B antibodies in your
blood plasma.

               Blood group B

If you belong to the blood
group B, you have B antigens
on the surface of your RBCs
 
and A antibodies in your
blood plasma.
Blood Antigens
Antibodies in Serum Genotypes
Group on RBCs

A A Anti-B AA or AO
B B Anti-A BB or BO
AB A and B Neither AB
O Neither Anti-A and anti-B OO
 ABO Routine Testing
Indirect/Reverse Typing

 Known antigen (cell) vs unknown antibody (patient’s

serum)

 Serum is combined with cells having known antigen

content in ratio 2:1 ratio

 Uses commercially prepared reagents containing saline

suspended A1 and B cells
Reaction Patterns for ABO
Blood Group Agglutination w/ Anti-A Agglutination w/ Anti-B

A + -

B - +

AB + +

O _ _
 ABO Routine Testing
Indirect/Reverse Typing

 Known antigen (cell) vs unknown antibody (patient’s

serum)

 Serum is combined with cells having known antigen

content in ratio 2:1 ratio

 Uses commercially prepared reagents containing saline

suspended A1 and B cells
Reaction Patterns for ABO
Blood Group Agglutination w/ A cells Agglutination w/ B cells

A _ +

B + _

AB _ _

O + +
 Slide Blood Typing
The slide is divided into halves.
On one side a drop of anti-A is added, this will attach to
and cause clumping of rbcs possessing the A antigen.
On the other side a drop of anti-B is added which will cause
clumping of rbcs with the B antigen.
A drop of rbcs is added to each side and mixed well with
the reagent.
The slide is tilted back and forth for one minute and
observed for agglutination (clumping) of the rbcs
Interpretation of Slide Typing
Testing with Anti-A Anti-Serum
If an rbc contains the A antigen
the red blood cells will be
agglutinated by anti-A, a
positive reaction.
If an rbc does not have the A
antigen there will be no
clumping, a negative reaction.
Interpretation of Slide Typing
Testing with Anti-B Anti-Serum
If an rbc contains the B antigen
the red blood cells will be
agglutinated by anti-B, a positive
reaction.
If an rbc does not have the B
antigen there will be no clumping
by anti-B, a negative reaction.
Slide Blood Typing Group A
An unknown rbc suspension is added to known anti-sera.
The left hand of the slide contains anti-A which reacts
with the unknown cell.
The right hand side contains anti-B which does not react
with the cell.
Slide Blood Typing Group B
An unknown rbc suspension is added to known anti-sera.
The left hand of the slide contains anti-A does not react
with the unknown cell.
The right hand side contains anti-B which reacts with the
cell.
Slide Blood Typing Group O
The left hand of the slide contains anti-A does
not react with the unknown cell.
The right hand side contains anti-B does not
react with the unknown cell.
Slide Blood Typing Group AB
The left hand of the slide contains anti-A which
reacts with the unknown cell.
The right hand side contains anti-B which reacts
with the unknown cell.
Summary of Slide Typing
Anti-A Anti-B Blood Group

NEG NEG O

POS NEG A

NEG POS B

POS POS AB
Example of Determining Genotype
Mom’s phenotype is group A, genotype AO
Dad’s phenotype is group B, genotype BO

B O
A AB 25% AO 25% (Group A)
O BO 25% (Group B) OO 25% (Group O)
Rh Blood Group
Another common way to group blood types is using Rh
antigens
Rh stands for ‘Rhesus’ because the Rh antigen was first
found in the Rhesus monkey
People with the Rh antigen are ‘Rh positive (Rh+)’, whereas
those without the antigen are ‘Rh negative (Rh -)’
Blood plasma does not normally contain anti-Rh
antibodies
Hemolytic Disease of the Newborn: Rh
Disease
• The most common problem with Rh incompatibility is during
pregnancy
• A small amount of blood from the fetus may contact the blood of the
mother through the placenta

If the fetus has Rh+ blood and the mother

has Rh- blood, then the mother will make
anti-Rh antibodies
However, contact between fetal and maternal
blood is greatest during childbirth, and thus
the firstborn baby is usually not affected
Treatment of HDN
• If a woman has Rh- and gives birth to a child, or if she
has a miscarriage or abortion, she is given an injection
of anti-Rh antibodies called anti-Rh gamma globulin or
RhoGAM to prevent HDN.

• The antibodies bind to the fetal Rh antigens and

inactivates them if they crossed the placenta during
birth, and the mother’s immune system does not
respond by producing antibodies.