ABO GROUPING CELL AND SERUM

GROUPING
SUBJECT HEMATOLOGY
(PRACTICAL)
SUBMITED TO HAFIZ IFTIKHAR
HUSSAIN
SUBMITTED BY NATASHA SALEEM (046)
M.TALHA (047)
ABDUL SAMAD(048)
NAIMA MUKHTAR(049)
MATEEN(050)
ABO SYSTEM
In ABO system there are four main types of blood groups :
A,B,AB and O identification of these four
blood groups is based on presence or absence of A and
/or B antigens on red cells.
Landsteiner rule If there is antigen on a red cell ,the
absence of it's corresponding antibody in the serum ( if
you're A you don't have anti-A
Blood group A :A antigens on the surface of RBCs and B
antibodies in blood plasma
Blood group B :B antigens on the surface of RBCs and A
antibodies in blood plasma
Blood group AB: Both A and B antigens on the surface of
RBCs and no A or B antibodies at all in blood plasma
Blood group O :Neither A or B antigens on the surface of
your RBCs but both A and B antibodies in blood plasma
HISTORY:KARL LANDSTEINER
Discovered the ABO blood group system in 1901.He and his
five co-workers began mixing each others red cells and serum
together and inadvertently performed the first forward and
reverse ABO groupings.
The forward grouping suggests the presence or absence of A
and B antigens in RBCs, whereas reverse grouping indicates
the presence or absences of anti-a and anti-b in serum
Agglutination reactions
Agglutination occurs when incompatible blood groups are
mixed together and this results in the clumping ( not
clotting) of blood.
For instance, if blood group A (which has A antigens on its
erythrocytes and B antibodies in its blood plasma) is
mixed with blood group B (which has B antigens on its
erythrocytes and A antibodies in its plasma), the A
antibodies would cause agglutination with the A antigens
on the red blood cells
Universal Donor and Recipient
UNIVERSAL DONOR GROUP O
➢ Neither A or B antigens
UNIVERSAL RECEIPIENT GROUP
AB
➢ Patient has no Anti A/Anti B present
Routine testing for ABO BLOOD GROUP
❑ Routine testing for determining the ABO GROUP
consist of testing the red blood cells with anti-A and
anti-B.
❑ It called forward typing
❑ Another test is perform by Checking antibodies in
plasma.
❑ It called reverse grouping.
❑ It is opposite to cell grouping.
Purpose of ABO grouping
❖ Blood typing is a method to tell what type of
blood you have.
❖ Blood typing is done so you can safely donate
your blood or receive a blood transfusion.
❖ It is also done to see if you have a substance
called Rh factor on the surface of your red blood
cells
ABO TYPING TECHNIQUES
• Slide test
❑ Slide testing imposes a greater risk
of exposure to infectious samples.
❑ Person should follow safety measures
detailed in the facility’s procedures
manual.
❑ Slide testing is not suitable for detection
of ABO antibodies in serum/
plasma.
Materials required for slide method
❑ Slides
❑ Antisera-A
❑ Antisera-B
❑ Antisera-D
❑ Blood specimen
❑ Disposable mixing sticks
Slide method
➢ Place 1 drop of anti-A on a clean, labeled
glass slide.
➢ Place 1 drop of anti-B on a separate
clean, labeled glass slide.
➢ Place 1 drop of antisera-D on a separate slide.
➢ Add a drop of blood on each drop of antisera.
➢ Mix with stirrer
➢ Gently tilt the slide continuously for
up to 2 minutes
Read, interpret, and record the results
of the reactions on all slides
Interpretation
❑ Strong agglutination of red cells in the presence of
any ABO typing reagent constitutes a positive
result.
❑ A smooth suspension of red cells at the end of 2
minutes is a negative result.
Advantages
1.Can be used in emergency and blood camps
for preliminary grouping
2. Easy to perform
3. Quick
Disadvantages
1.Not reliable for weak reactions
2. Serum testing cannot be performed
3. The test mixture tends to dry fast
4.drying causes aggregation of cells which can
be interpreted as agglutination
5. Less sensitive than tube technique
Test Tube Method (Gold standard)
❖ Allows longer incubation of antigen and antibody
mixture
without drying
❖ Tubes can be centrifuged to enhance reaction
❖ Can detect weaker antigen / antibody
❖ Two steps in ABO grouping Cell grouping
❖ Forward grouping – Tests the patients red cells
with known Anti-A & Anti- B to determine the antigen
expressed Serum grouping
❖ Reverse grouping – Test the patients serum with
known A & B cells to determine the presence of
antibody
Material required
• Test tube
• Antisera-A
• Antisera-B
• Antisera-D
• Normal saline
• Dropper
• 5% cell suspension
• Serum
• Centrifuge machine
CELL GROUPING ( Forward grouping)
❑ Place 1 drop of anti-A in a clean, labeled
test tube
❑ Place 1 drop of anti-B in a clean, labeled
Tube
❑ Add to each tube 1 drop of a 5% suspension of the
red
cells to be tested.
❑ Mix the contents of tube gently and centrifuge them
At 1500 rpm for 1 minute
Read, interpret, and record the test
results.
Interpretation
❑ Agglutination of tested red cells and either
hemolysis
or agglutination in tests with serum constitute
positive
test results.
❑ A smooth cell suspension after
resuspension of the cell
button is a negative test result
5% cell suspension
❑ place a 2-3 drops of blood into tube
❑ Fill the tube with saline
❑ Centrifuge at 3000 rpm for 2 minute
❑ Discard the serum or plasma
❑ Again fill tube with saline and repeat this process
at
least three time’
❑ And then to make 5% cell suspension add 1 drop
of
packed red blood cell to 19 drop of saline.
SERUM GROUPING ( REVERSE GROUPING)
1.Prepare 2-5% suspension of cells A,B
2. Label three tubes A cells, B cells
3. Place two drops of serum in each tube
4. Add one drop of cell suspension ( A cell to A tube, B
cell
to B tube
5. Reverse Grouping Centrifuge at 1000 rpm for 1 min
6. Centrifuge & record the results
Advantages
1.Easy to perform
2.accurate and more sensitive than slide
technique
3. The cell mixture can be incubated for a long
time without drying.
4.The centrifugation used enhances the reaction
and hence even weak antigens / antibodies can
be detected
5.uses smaller quantity of reagents
6.clean and more hygienic
Disadvantages
1.Requires Labeling and manipulation of tubes
2. Always use thoroughly cleaned test tubes for test
3. While reading the result ,shake very gently to dilodge
the button.
4. Use correct speed and time centrifugation to avoid
erroneous results.
Reference
https://youtu.be/H6w-BRSgfMg
https://youtu.be/y6VIMkUO3OA
book: Human blood groups GEOFF DANIELS(Second
Edition)
book: AABB technical manual 15TH edition