DETERMINATION OF
BLOOD GROUPS
Aim of the experiment
To determine the blood group of given blood sample.
Apparatus
Blood grouping tile
Anti A serum(monoclonal anti -A antibodies)
Anti B serum(monoclonal anti-B antibodies)
Anti D serum(monoclonal anti-Rh(D)antibodies)
Anticoagulant (1%sodium citrate in normal saline)
Sterile Cotton swab
Spirit
lancet
3 tooth picks
Marker pen
Principle
The surfaces of red cell membrane contain a variety of
genetically determined antigens , called ISO ANTIGENS
or AGGLUTINOGENS.
To determine the blood group of a person ,the red cells
are made to react commercially available antisera known
agglutinins .
The slide is examined under microscope to detect the
presence or absence of clumping and hemolysis of red
cells,result of antigen antibody reaction.
Procedure
Take a clean blood grouping tile and make three
columns with a marker pen
Name each column as A , B , D with marker pen
Place a drop each of anticoagulant (sodium citrate
solution) into each column
Wipe the ring finger with cotton spirit swab
Allow the spirit to dry
Make a gentle prick with needle
Place one drop of blood in three columns ,with
anticoagulant ,without finger touching the tile
Place a drop of Anti A serum in A column
Place a drop of Anti B serum in B column
Place a drop of Anti D serum in D column
With the help of three separate tooth picks, mix the
serum and blood drop .
Wait for a minute and record for presence or absence
of agglutination in each column
Interpret the result
Observation
Report
The blood group of given blood sample is B Positive.
Questions:
1Q. What is a blood group system? What is the
physiological and genetic basis of blood
grouping?
The surfaces of human red cells contain a variety of
genetically determined glycolipids and glycoproteins
that act as antigens.
The plasma contains antibodies that can react with these
antigens when the two are mixed. Since the red cell
antigens cause agglutination of RBCs in the
presence of suitable antibodies, they are also called
agglutinogens and the antibodies in the plasma are
called agglutinins.
2Q. What are the precautions taken for
determination of ABO blood groups?
1) The columns must be labelled correctly.
2) One Tooth pick should be used for one column
only.
3) Don’t use same tooth pick for other column.
4) Use equal amount of blood and anti sera drops in
each column.
3Q. How will you confirm the clumping of red
cells on a glass slide?
Before the slide dries up, under low power objective of
microscope agglutination is to be noted.
4Q. What is the mode of inheritance of blood
groups?
1)Presence of blood group antigen is co dominant
characteristic.
2)Antigen is present in the phenotype regardless of
the genotype.
5Q. What is Landsteiner’s law and what are the
exceptions to this law?
Landsteiner’s law(1990)- 2 major components:
If an agglutinogen is present on RBC membrane, the
corresponding agglutinin must be absent in the plasma.
If an agglutinogen is absent on RBC membrane, the
corresponding agglutinin must be present in the
plasma.
Exception to 2nd part of this law-Rh system.
6Q. What is a universal donor and a universal
recipient?
Universal donor – O negative.because they don’t have
either A or B antigens on their red cells ,theoratically
their blood can be given to all the 4 blood groups.
Universal recipient – AB positive,becoz they don’t have
circulating agglutinins in their plasma and therefore
receive blood from any type.
7Q. What is cross matching of blood and what is
its clinical significance?
Two types of cross matching:
Major cross matching – the cells of the donor are
directly matched against the plasma of the recipient. It
is important to ensure that antibodies present in the
recipient’s plasma do not harm the donor’s red cells.
Minor cross matching – The donor’s plasma is
checked against the RBCs of the recipient. It is called
minor cross matching . A small volume of donor’s
plasma is diluted in large volume of recipient’s plasma.
It is not so important
8Q. What is Rh incompatibility and how does it differ from
ABO incompatibility?
When an Rh negative person receives blood from an Rh positive
person , there is a reaction and it is called
Rh incompatibility.
Rh incompatibility differs from ABO incompatibility in terms of
speed with which transfusion reaction occurs.
Transfusion reaction doesn’t occur in first transfusion. In 2 nd
transfusion , reaction occurs due to preformed antibodies.
In ABO incompatibility, transfusion reaction occurs immediately.
9Q .What is erythroblastosis fetalis and how do you prevent it?
Hemolytic disease of the newborn due to Rh incompatibility.
When Rh negative mother carries Rh positive fetus, first time
nothing occurs.
A small amount of blood leaking into maternal circulation at
the time of delivery induces formation of anti Rh agglutinins
in the mother.
In next pregnancies, the mother’s agglutinin crosses the
placenta to the fetus and causes hemolysis.
Prevention – administration of a single dose of anti Rh
antibodies in the form of Rh immunoglobulin within 24 hrs of
first delivery.
10.Q Name few major blood group systems and minor blood
group systems?
Major blood group systems –
A) ABO blood group system.
B) Rh system
C) MN system
Minor blood group systems –
A) Lewis system
B) Kell system
C)Duffy system
