Blood grouping for DMLT

1. BLOOD GROUPING
PREPARED BY PUNAM SAHOO

2. INTRODUCTION
• The ABO Blood group represent the antigen present on red blood cells of each
group and whenever antigen A or B is absent on the erythrocytes the
corresponding antibody is found in the serum .
• These antigens may be proteins, carbohydrates, glycoproteins depending on the
blood group system .
• There are nearly 300 blood group system so far discovered. Of these ABO and Rh
are major, universally adapted and medically important of all the blood groups.
• It is most important that blood donor and recipient are grouped correctly before
transfusion .

3. ABO BLOOD GROUP SYSTEM
Karl Landsteiner, an Austrian scientist discovered the ABO blood
group system in the year 1900.
During the blood transfusion, the two most important group systems
examined are the ABO-system and the Rhesus system.
The ABO blood group system consists of 4 types of blood group – A,
B, AB, and O and is mainly based on the antigens and antibodies on
red blood cells and in the plasma.
Both antigens and antibodies are protein molecules in which antigens
are present on the surface of Red Blood Cells and antibodies are
present in the plasma which is involved in defending mechanisms.
Rh System:-The presence of Rh antigen on the RBC surface called
Rh positive group and absence of Rh antigen called Rh negative
group.

5. BLOOD GROUPING METHOD
PRINCIPLE:-
ABO blood grouping methods are based on process of hemagglutination when red
cell antigen react with its corresponding antibodies the clumping (agglutination ) is
seen.
Blood grouping occurred by 2 methods.
1. FORWARDDIRECT BLOODGROUPING:-
 Direct blood grouping also known as cell grouping.
 In this test person’s red cells are tested with reagent anti —A and anti —B sera.
 It can be performed by two methods
A. Test tube method
B. Slide method

6. TEST TUBE MATHOD
EQUIPMENT'S & REAGENT — Centrifuge machine, Test tube, Dropper, Anti- A
Serum, Anti- B Serum.
PROCEDURE:-
Take two test tube.
Label one tube with 'anti-A and the second 'anti-B'
 Add one drop of anti —A serum into the labelled anti-A' test tube.
 Add one drop of anti —B serum into the labelled 'anti-B' test tube.
 Add one drop of 2-5 % red cell suspension into both tubes.
 Mix well the anti serum and cells suspension into both tubes and gentle shaking.
 Leave the tubes at room temperature for 5 minutes and centrifuge at 1500 rpm for 60 seconds.
 Read the result by gently topping the base or bottom of tube for agglutination.

7. INTERPRETATION :
Observe the slide for 2 minutes.
Positive — if agglutination occur.
Negative -- if no agglutination occur.

8. SLIDE METHOD
EQUIPMENT'S & REAGENT –
Centrifuge machine, Slides, Anti-A Sera, Anti —B Sera, Normal Saline .Blood specimen —Venous
or capillary blood with no preservative.
PROCEDURE:-
 Take a dry and clean glass slide.
 Make a ceramic ring on the half of slide.
 Label one ring as Anti-A and other ring as Anti-B.
 Add one drop of anti —A sera on the ring labelled anti-A. Add one drop of anti —B sera on the
ring labelled as anti-B.
 Add one drop of red cell suspension on both rings. Mix well cell and serum mixture using
separate application stick.
 Observe the slide for any agglutination by tilt the slide back and forth.

9. INTERPRETATION
Observe the slide for 2 minutes.
Positive + if agglutination occur.
Negative — if no agglutination occur.

10. REVERSE BLOOD GROUPING
 This is also known as indirect blood grouping or serum grouping.
 It is opposite to direct method of grouping individual scrum is tested with known group A cells and B cells.
 The reverse blood grouping can be performed in two methods: Tube and Slide method. The Tube method is
preferred to slide method.
TEST TUBE METHOD:-
 Label two test tubes as A and B.
 Add two drops of serum to be tested in each tube.
 Add one drop each of A and B cells suspension to the corresponding test tubes.
 Mix well and centrifuge both tubes at 1000 rpm for 1 minute.
 Gently remove the tubes and completely resuspend cells and examine macroscopically for agglutination and if
negative, microscopically.
 Record the reactions and interpret the results.

11. SLIDE METHOD
PROCEDURE
Mark a clean slide into two halves, labeling the left and right side slide as A and B.
Add a drop of serum to be tested on both sides.
Add one drop each of A and B cells suspension (20%) to the corresponding sides.
Using a clean applicator stick, mix the serum and cell suspension on both sides
separately and spread into a smooth round circle.
Rock the slide gently for 2 minutes and look for agglutination.
Record the reactions and interpret the results.

12. Interpretation:-
 If agglutination is observed with A cells only, then the patient’s
blood group is B.
 If agglutination is observed with B cells only, then the patient’s
blood group is A

13. Rh BLOOD TYPING
INTRODUCTION:-
Human red blood cells are classified as Rh positive or Rh negative depend upon
the presence or absence of the D-Antigen or Rh antigen.
 Rh typing is also known as Rh blood grouping test. o Rh typing is done along
with ABO blood grouping.
PRINCIPLE:
Rh typing is based on the principle of agglutination.
 The red blood cells of individual possess Rh antigen (also called D antigen) will
agglutinate or clump with anti —D anti serum .
if agglutination occurs it is called Rh positive and no agglutination consider Rh
negative.

14. METHODS
Usually two methods are performed for Rh typing—
1. A. Tube method
2. B. Slide Method
EQUIPMENTS & REAGENTS:-
1. Test tube or slide
2. Dropper / Pipette.
3. Anti-D serum.
4. Centrifuge machine.
5. Blood sample

15. Tube method
PROCEDURE
First of all prepared 2-5 % suspension of red blood cell with the mixing of normal saline. Take
clean and dry test tubes.
 Add one drop of anti —D serum in each test tube. Add one drop of cell suspension in tube.
Mix well anti —D serum and suspension.
Centrifuge the tubes at 1500 rpm for 60 seconds incubate at room temperature for a minute.
 Observe the test tube for agglutination reactions.
INTERPRETATION:-
Presence of agglutination consider Rh positive
No agglutination is consider Rh negative.

16. ADVANTAGE OF TUBE METHOD
• Tube method is more sensitive then slide method Long incubation
without drying tube content is possible with tube method.
• Small volume of reagent required for this method
• Interpretation is easy with simplicity of reading

17. SLIDE METHOD
PROCEDURE:-
Take clean and dry glass slide.
Put one drop of anti —D sera on slide.
Add one drop of 5 % suspension of red blood cells by pipette
Mix well cell and serum using applicator stick.
 Observe for agglutination for Two minute.
INTERPRETATION:-
No agglutination —Rh Negative.
Agglutination Seen —Rh Positive.

18. Significance of Rh:-
If the mother is Rh negative and father is Rh positive with a fetus who
is Rh positive.
Here body will intake antibodies against the fetus's blood
when small amount of the baby's blood mixes with maternal blood.
The mother's body makes antibodies to Rh antigens The mother
antibodies may cross placenta and breaks the fetus red blood cell this
can cause Rh disease also known as hemolytic disease of new born.

20. CONCLUSION
The ABO and Rh blood group system is of prime importance as it helps in differentiating blood.
The blood of individuals who require blood transfusions is matched with the donor’s blood before
carrying out a blood transfusion.
 Matching a blood group is essential because if the blood is not compatible, the recipient may
suffer severe consequences, even death. When a recipient receives incompatible blood, the
antibodies attack the red blood cells resulting in agglutination in the blood vessels, bursting of
cells, and hemolysis, which can sometimes be fatal.
Pregnant women are also required to undergo a blood group test to ascertain the Rh factor of the
mother and the baby. If the mother is Rh D negative and the baby is Rh D positive, it could lead to
complications in pregnancy. With the advent of blood grouping systems, it has been possible to
prevent unnecessary complications in blood recipients and pregnant women.