ABO

INCOMPATIBILITIES

Dr. Ni Kadek Mulyantari, Sp.PK(K)

(Department of Clinical Pathology Medical
Faculty Udayana University/Sanglah
General Hospital Denpasar)
 INTRODUCTION
Blood transfusion is the oldest and most
frequently tissue transplantation
The first human blood group ABO, was
describe in 1900, and with the anticoagulant
(1914) we can store the blood for
transfusion, first blood bank hospital was
establish in 1936
 INTRODUCTION
There are no absolute indications and few
contraindications to blood transfusion
All decisions about blood transfusion must be
driven by the clinical considerations such as the
cause of bleeding, amount and rate of blood
loss, underlaying diseases, risk of future
bleeding & extent of physiologic compensation
 INTRODUCTION

Activities in blood centers and transfusion services. Product quality is the focus of
blood centers; clinical outcome is the focus of transfusion services.
 BLOOD DONATION
May be allogeneic or autologous
FDA establish criteria for acceptability of blood
donors. This include measures to protect donors
health and measures to protect the resipients
Giving blood:
Single blood donation is 450ml  blood components
Apheresis  less effect on blood volume so a greater
amount of intended component can be collected
from one donor.
 Donor testing
Blood donations are tested for markers of infectious
diseases that may be transmitted.
HBs Ag
Anti-HBc CURRENT INFECTIOUS
DISEASE TESTING
Anti-HCV
HIV antibody
Serologic test for syphillis
HIV , HCV, HBV genome (NAT)
The most importance in reducing transfusion
transmitted diseases is medical histories
 Blood Types
Categorized according to antigens and antibody
on red blood cells
Type A A antigens, antibody B
Type B:B antigens, antibody A
Type O: no antigens, antibody A&B (universal donor)
Type AB A and B antigens, no antibody (universal recipient)
D antigen, third antigen; may be present on the red
blood cells
• a. Rh factor positive: D antigen is present
• b. Rh factor negative: D antigen is not present
 COMPATIBILITY TESTING
All steps in the identification and testing of a
potential transfusion recipient and donor
blood before transfusion in an attempt to
provide a blood product that survives in vivo
and provides its therapeutic effect in the
recipient
Process of compatibility testing
Pre-transfusion testing (WHO)
It is essential that all blood is tested before transfusion in order to:
• Ensure that transfused red cells are compatible with antibodies in the
recipient’s plasma
• Avoid stimulating the production of new red cell antibodies in the
recipient, particularly anti-RhD.

All pre-transfusion test procedures should provide the following

information about both the units of blood and the patient:
• ABO group
• RhD type
• Presence of red cell antibodies that could cause haemolysis in the
recipient.
The laboratory performs:
• Patient’s ABO and RhD type
• Direct compatibility test or crossmatch.

These procedures normally take about 1 hour

to complete. Shortened procedures are
possible, but may fail to detect some
incompatibilities.
 Method for ABO Blood group testing
• Slide test / glass slide / white porcelain tile
• Tube test
• Microwell plate or microplate test.
• Column technique (sephadex gel)
• Solid phase tests.
 Slide test / glass slide / white porcelain tile

Procedure
1. The test can be performed either on glass slides or on ceramic
tiles.
2. Place one drop of anti-A and one drop of anti-B sera on two
previously labelled slides.
3. Add one drop of blood (preferably 20% red cell suspension) on
each slide.
4. Mix properly by a clean glass stick or the corner of another slide.
5. Rock the slides in order to mix the cells and sera and leave at
room temperature for 2 minutes.
6. Record the results.
Slide test / glass slide / white porcelain tile
Interpretation of results of
ABO grouping
 Tube test

Cell grouping (forward grouping)

1. Prepare 5% red cell suspension (tomato colour) in normal saline.
2. Add 1 drop of anti-A in the tube labelled A, anti-B in the tube labeled B
and anti-AB in the tube labelled AB.
3. Add 1 drop of the cell suspension in each tube.
4. Mix properly, incubate the mixture at room temperature (RT) for 5-10
minutes and then centrifuge at 1000 rpm for 1 minute.
5. If no haemolysis is observed in the supernatant, disperse the cell button.
6. Check for agglutination. If no clump is seen by naked eyes, examine
under microscope for weak agglutination.
7. Record the results.
 Tube test
Serum grouping ( Reverse grouping)
1. The serum of the donor/patient is tested against known cells of group A, B and O. These
cells are either prepared in the lab by pooling or can be acquired from manufacturers.
2. Arrange three test tubes and label them A, B and O.
3. Place 2 drops of the serum to be tested in each tube.
4. Add 1 drop of A group cells to the tube A, B group cells to tube B and O group cells to the
tube labelled O.
5. Shake the contents gently. Incubate at RT for 5-10 minutes and centrifuge at 1000 rpm
for 1 minute.
6. If the supernatant shows no signs of haemolysis, disperse the cell button and observe for
agglutination.
7. If no agglutination is observed by naked eyes, examine under microscope.
8. Record the results.
Tube test
Interpretation ofTube test result
 Grades of agglutination
The agglutination results are graded from 1+ to 4+.
The American Association of Blood Banks (AABB)
recommends the following grading system:
• 4+ = One solid aggregate of red cells
• 3+ = Several large aggregates
• 2+ = Medium sized aggregates with a clear background
• 1+ = Small aggregates with a turbid background giving granular
appearance.
• Weak (w) = Tiny aggregates are seen only under microscope
• Negative = All cells are free.
ABO Blood group testing
Crossmatching
Crossmatch is usually performed before blood is infused.
The cross-matching is of two types: major cross-match and minor
crossmatch.

Major cross-match
When the donor’s red cells and the patient’s serum is mixed and
tested, it
is called major cross-match: Donor’s cells + Patient’s serum.

Minor cross-match
Donor’s serum + Patient’s cells
 Procedure of cross-matching
Saline Phase
This technique is designed to detect IgM antibodies of ABO system.
• Label two test tubes, one A for major and one B for minor cross-match.
• Place 2 drops of patient’s serum in the tube labelled A and 2 drops of donor’s serum in
the tube labelled B.
• Add 1 drop of 2-5% (tomato coloured) cell suspension of the donor in tube A and 1
drop of patient’s cells in tube B.
• Mix and incubate at RT for 10 minutes.
• Centrifuge at 1000 rpm for 1-2 minutes.
• Place 1 drop from each tube A and B on two separate glass slides and examine under
microscope.
• Put a drop of saline on each slide to disperse the rouleaux.
• Record the results.
 Albumin phase
• Add 1-2 drops of 22% bovine albumin to the serum
cell mixture.
• Incubate at 37°C for 30 minutes.
• Centrifuge at 1000 rpm for 1 minute.
• Haemolysis or agglutination at this stage indicates
presence of weak reacting (IgG) antibodies.
• Wash the cells three times with normal saline and
discard the supernatant
• completely every time.
 AHG/ IAT phase
• Add 2 drops of AHG serum.
• Centrifuge at 1000 rpm for 1 minute.
• Dislodge the button gently and examine for
agglutination.
• Haemolysis or agglutination at this stage
indicates presence of IgG
• antibodies.
Diagrammatic presentation of ABO
compatibility
Blood Components
Prepared from Whole blood collection or apheresis
Whole blood is separated by differential centrifugation
Red Blood Cells (RBC’s)
Platelets
Plasma
Cryoprecipitate
Others
Others include Plasma proteins—IV Ig, Coagulation
Factors, albumin, Anti-D, Growth Factors, Colloid
volume expanders
Apheresis may also used to collect blood components
RED CELL COMPONENTS
In red cell transfusion, there must be ABO and RhD
compatibility between the donor’s red cells and the
recipient’s plasma.
1. Group O individuals can receive blood from group O donors only
2. Group A individuals can receive blood from group A and O donors
3. Group B individuals can receive blood from group B and O donors
4. Group AB individuals can receive blood from AB donors, and also from
group A, B and O donors

Note: Red cell concentrates, from which the plasma has

been removed, are preferable when non-group specific
blood is being transfused.
PLASMA AND COMPONENTS CONTAINING
PLASMA
In plasma transfusion, group AB plasma can be given to
patient of any ABO group because it contains neither
anti-A nor anti-B antibody.

1 Group AB plasma (no antibodies) can be given to any ABO group patients
2 Group A plasma (anti-B) can be given to group O and A patients
3 Group B plasma (anti-A) can be given to group O and B patients
4 Group O plasma (anti-A + anti-B) can be given to group O patients only
 Plasma

O O A B AB
A A AB
B B AB
AB AB
 ABO incompatibility: haemolytic reactions
• Anti-A or anti-B recipient antibodies are almost always capable of
causing rapid destruction (haemolysis) of incompatible transfused
red cells as soon as they enter the circulation.
• A red cell transfusion that is not tested for compatibility carries a
high risk of causing an acute haemolytic reaction. Similarly, if blood
is given to the wrong patient, it may be incompatible.
• Typically, at least one third of unmatched transfusions will be ABO
incompatible and at least 10% of these will lead to severe or fatal
reactions.
• In some circumstances, it is also important that the donor’s
antibodies are compatible with the patient’s red cells. It is not
always essential, however, to give blood of the same ABO group.
 Time limits for infusion

There is a risk of bacterial proliferation or loss of function in blood

products, once they have been removed from the correct storage
conditions
 Warming blood
There is no evidence that warming blood is beneficial to the
patient when infusion is slow.
At infusion rates greater than 100 ml/minute, cold blood may be a contributing factor in
cardiac arrest. However, keeping the patient warm is probably more important than
warming the infused blood.

Warmed blood is most commonly required in:

Large volume rapid transfusions:
Adults: greater than 50 ml/kg/hour
Children: greater than 15 ml/kg/hour
Exchange transfusion in infants
Patients with clinically significant cold agglutinins.
Blood should only be warmed in a blood warmer.
Complications of transfusion
Refference
THANK
YOU