Blood Typing

Blood
• Blood is a constantly circulating fluid providing the body
with nutrition, oxygen, and waste removal. Blood is mostly
liquid, with numerous cells and proteins suspended in it, making
blood "thicker" than pure water
• Each red blood cell is about one-third hemoglobin, by
volume. Plasma is about 92% water, with plasma proteins as
the most abundant solutes.
• The main plasma protein groups are albumins, globulins, and
fibrinogens. The primary blood gasses are oxygen, carbon
dioxide, and nitrogen.
The human ABO blood groups were discovered by Austrian-born
American biologist Karl Landsteiner in 1901.
What is an antigen?
• An antigen is any substance that causes your immune
system to produce antibodies against it.
Types of Antigen
• Antigen A
• Antigen B
What is an antibody?
• An antibody is a protein produced by the body's immune
system when it detects harmful substances, called
antigens.
The antigens and antibodies combine by a process called agglutination.
• Your blood sample is mixed with antibodies against type A and B
blood. Then, the sample is checked to see whether the blood cells
stick together
• If blood cells stick together, it means the blood reacted with one of the
antibodies.
• Monoclonal sera ANTI-A, ANTI-B and ANTI-AB are specific IgM
immunoglobulin which are directed against the human red blood
cell antigens A, B and AB respectively.
• Antigens ( agglutinogens )starts appearing during 6 weeks of the fetal
life. Antigens are also present in the many organs i.e pancreas, salivary
glands, liver, lungs and kidneys
• Antigens enter in the body through the respiratory and digestive
system along with Bactria
• Antibodies ( agglutinins) starts appearing after 2- 3 weeks after the
birth
Antigen H
• This is the precursor of the RBCs
• They need enzymes to make antigen H as we know to produce the
enzymes and proteins, we need genes
• There is no nucleus in RBC so how they produced …. They are made
on the surface of the erythropoietic precursors cells ( these
erythropoietic precursors cells have nucleus
Where the H Substance comes from
 there is synthesis of oligosaccharides ( short sugar chains ) all RBC
have oligosaccharides precursor molecules ( millions pf such
molecules)
• These oligosaccharides are produced by the erythropoietin precursors
cells they are modified by the chromosome no 19 ( which produces
fructose transferase enzyme)
• If this enzyme express, it will attach the fructose molecule with the
chain of the oligosaccharides
• Now the oligosaccharides molecule with the fructose is called H
substance
• This happens in all and every individual either you have blood group A
or B or some other
• In blood group O they do not have transferase A or B ( there is zero
conversion)
Antigen A
• There is additional activity of the chromosome no 9 which releases
transferase A ( transfer N- acetyl galactosamine NAGA)
• Now oligosaccharides with fructose and NAGA is called substance A
• W hen H substance is added with the N-acetyl galactosamine then H
substance is converted into A antigen
Antigen B
• Activate chromosomal 9 due to slight variant in the gene locus so, it
make different type of the transferase not NAGA
• It is transferring only galactose ( galactose transferase)
• When H substance added with the galactose then H substances is
converted into B antigen
ANTIGEN ANTIBODY BLOOD TYPE

A B A •AMTIGEN
•ANTIBODY
B A B •BLOOD TYPE

X AB O

AB X AB
RH Blood Group System
• RH antigen is protein and peptide chain which passes through the red
blood cells 12 times
• About 50 Rh antigens has been discovered but 5 are more important
-D antigen factor
-C /c antigen
-E/e antigen
Rh Factor
• The Rh factor is an
inherited protein that can
be found on the surface of
the red blood cell. If your
blood type is positive, then
your blood cells have the Rh
protein. If your blood type is
negative, then your blood
cells lack the Rh protein.
• Transmembrane proteins
which transports the
ammonia and carbon dioxide
• 85 % population is RH
positive
• While 15 % is RH negative
Blood group with RH antigen
• Antigen A + antigen D ------------- A positive
• Antigen B + antigen D ------------- B positive
• Antigen A +No antigen D---------- A negative
• Antigen B + No antigen D---------- B negative
• Antigen A,B+ No antigen D---------- AB negative
• Antigen A,B+ antigen D---------- AB positive
• H antigen ( not A not B)+ no D antigen--------- O negative
• H antigen ( not A not B)+ D antigen--------- O positive
ABO vs RH system
• RH antigens are protein in nature while the ABO antigens are carbohydrate in
nature they are not protein in nature
• Antibodies against antigen A and B are produced by naturally spontaneously while
the antibodies against the RH antigen are not produced naturally spontaneously
• RH antigens are not widely found in the diet and microbes while ABO antigens
are present
• RH system can stimulate T cells while ABO cannot stimulate the T cells
• If there is incompatibility in ABO system there is no or less hemorrhages in fetus
but if incompatibility is due to RH there is severe damage and hemorrhages
• ABO antigens are present on RBC as well as others while RH antigens are
expressed only on the RBC cells
• Anti RH and anti ABO damage the cell in different way
• Gene which produces the RH factor primarily present on the short arm of
chromosome no. 1 basically it consist of two genes ( RHD and RHd )
RHD = means gene is present and fully expressed
RHd = means gene is absent or dysfunctional
• Genes for ABO is present on chromosome no. 9
• AB antigens are express after birth but RH antigens are expressed intra fetal life
• RH positive person may be homozygous or heterozygous while RH negative
person only homozygous
Antisera A
Tint blue in color
Obtained from B blood group
Contain alpha agglutinins
Antisera B
Tint yellow in color
Obtained from the A blood group
Contained Beta agglutinins
Antisera B
Contains RH agglutinins
Obtained from O blood group
White in color
 Procedure
a. Take clean glass slide and divide into three equal parts a, b , c.
b. Mark one part for A one for B and one for D antisera.
c. With the help of the lancet prick on the finger to ooze out the blood.
d. Place one drop of blood on three divided parts of the slide.
e. Add one drop of antisera A to a side.
f. Add one drop of antisera B to b side.
g. Add one drop of antisera D to c side.
h. Mix them with the help of toothpicks.
i. Use different tooth pics for each mixing.
j. Spread the mixture
k. Observe the agglutination with illuminator.
Antisera A Antisera B Antisera D BLOOD GROUP

✓ ✓ ✓ AB POSITIVE

✓ ✓ X AB NEGATIVE

✓ X X A NEGATIVE

✓ X ✓ A POSITIVE

X ✓ ✓ B POSITIVE

X ✓ X B NEGATIVE

X X X O NEGATIVE

X X ✓ O POSITIVE
Universal donor

Universal recipient

AB
Complications of
mismatched
• Jaundice
• Cardiac shock
• Renal shutdown
Erythroblastosis fetalis
• Disease in the newborn and fetus
• Abnormal hemolysis of the RBCs
• However, at the time of parturition (delivery of the child), the Rh antigen
from fetal blood may leak into mother’s blood because of placental
detachment. During postpartum period, i.e. within a month after delivery,
the mother develops Rh antibody in her blood.
• When the mother conceives for the second time and if the fetus happens to
be Rh positive again, the Rh antibody from mother’s blood crosses
placental barrier and enters the fetal blood.
• Thus, the Rh antigen cannot cross the placental barrier, whereas Rh
antibody can cross
• Rh antibody which enters the fetus causes agglutination of fetal RBCs resulting in
hemolysis. Severe hemolysis in the fetus causes jaundice.
• To compensate the hemolysis of a greater number of RBCs, there is rapid
production of RBCs, not only from bone marrow, but also from spleen and liver
• Now, many large and immature cells in proerythroblast stage are released into
circulation. Because of this, the disease is called erythroblastosis fetalis.
Ultimately
1. Severe anemia
2. Hydrops fetalis
3. Kernicterus.