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Transfusion Medicine Quality Manual
Standard Operating Procedure for ABO Grouping
Tube Method
Standard Operating Procedure for
ABO Grouping Tube Method
Provincial Blood
Coordinating Program
1.0
1.1 Each recipient blood sample for compatibility testing shall be tested for
ABO group.
1.2 The results of the red cell and plasma test must agree. Current and previous
results should be compared to identify any ABO discrepancy.
1.3 All blood group discrepancy shall be resolved and the resolution
documented before issuing red cells. If transfusion is necessary before
resolving the ABO discrepancy the recipient should receive group O red
cells and AB plasma products.
1.4 Blood group testing for A, B and D antigens only is required for:
1.4.1 Infants less than 4 months of age; and
1.4.2 Confirmation of the ABO group on donor units;
2.0
Patient Identification and Specimen Labeling. Available at:
http://www.health.gov.nl.ca/health/bloodservices/pdf/patient_id_and_specimen_la
beling.pdf
Determining Specimen Suitability. Available at:
http://www.health.gov.nl.ca/health/bloodservices/pdf/determining_specimen_suit
ability.pdf
Patient History Check. Available at:
http://www.health.gov.nl.ca/health/bloodservices/pdf/patient_history_check.pdf
Preparation of Red Cell Suspensions. Available at:
http://www.health.gov.nl.ca/health/bloodservices/pdf/preparation_of_red_cell_sus
pensions.pdf
Quality Control of Reagents and Antisera. Available at:
http://www.health.gov.nl.ca/health/bloodservices/pdf/quality_control_of_reagents
and_antisera_ver1.pdf
_______________________________________________________________________
This document may be incorporated into each Regional Policy/Procedure Manual.
NL2010.-008 –TMQ
Version: 3.0
Effective Date: 2015-04-03
Page 2 of 11
Standard Operating Procedure for
ABO Grouping Tube Method
Provincial Blood
Coordinating Program
Resolving ABO & Rh Blood Group Discrepancies. Available at:
http://www.health.gov.nl.ca/health/bloodservices/pdf/resolving_abo_rh_blood_gr
oup_discrepancies.pdf
3.0
3.1 All Transfusion Medicine Laboratory Technologists who are responsible for
testing and reporting transfusion medicine samples.
4.0
4.1 ABO blood groups are determined by phenotyping the recipient’s red cells
for the presence or absence of A and B antigens on red cells and by testing
the recipient’s plasma for the presence or absence of anti-A and anti-B.
4.2 ABO typing and ABO compatibility testing is the foundation of
pretransfusion testing.
4.3 Transfusion of ABO incompatible blood can be associated with acute
intravascular hemolysis, renal failure and death.
4.4 Routine tests to determine the ABO group consists of testing red cells with
anti-A and anti-B (forward group) and testing the plasma with A and B red
1
cells (reverse group).
4.5 Hemolysis is interpreted as a positive result.
4.6 Pre-washing the red cells is not necessary, however, if a discrepancy is
detected the cells should be washed and the tests repeated.
4.7 Washed red cells may reduce false positive results associated with rouleaux
or autoantibodies.
4.8 Positive reactions in the forward group characteristically demonstrate a
grade three (3) to grade four (4) agglutination. Reactions in the reverse
grouping (serum and reagent red cells) are often weaker.
4.9 Reactions in the forward group that demonstrate less than grade 2 reactions
should be further investigated.
_______________________________________________________________________
This document may be incorporated into each Regional Policy/Procedure Manual.
NL2010.-008 –TMQ
Version: 3.0
Effective Date: 2015-04-03
Page 3 of 11
Standard Operating Procedure for
ABO Grouping Tube Method
Provincial Blood
Coordinating Program
4.10 The serum tests may be incubated at room temperature for 5 to 15 minutes
to enhance weak reactions.
4.11 A mixed field reaction maybe detected if a recipient has been transfused
with ABO compatible red cells other than that of their own ABO group.
4.12 False positive or negative results can be caused by variables such as:
4.12.1 Improper technique
4.12.2 Contaminated materials
4.12.3 Omission of reagents or anti-sera
4.12.4 Delays in testing
4.12.5 Inadequate incubation time and temperature
4.12.6 Inappropriate centrifugation
4.12.7 Inappropriate or prolonged storage of red cells.
4.13 If performing the ABO grouping by automation follow manufacturer’s
instructions for operation and resulting.
5.0
5.1 Quality Control
5.1.1 All reagents shall be used and controlled according to the
manufacturer’s written instructions.
5.1.2 All anti-sera must be visually inspected for contamination such as
discoloration, cloudiness, turbidity and/or particulate matter.
5.1.3 All reagent red cells must be visually inspected for hemolysis and/or
discoloration.
5.1.4 The results of the visual inspection, reagent lot number, expiry date,
date of the inspection and the individual performing the inspection
must be documented.
5.1.5 The expiry date should be checked on each reagent used. Do not use
reagents beyond expiry date.
_______________________________________________________________________
This document may be incorporated into each Regional Policy/Procedure Manual.
NL2010.-008 –TMQ
Version: 3.0
Effective Date: 2015-04-03
Page 4 of 11
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