9 - ABO Discrpancy

ABO Discrepancies
Basic ABO Grouping

ABO group grouping Reverse Grouping Forward
B cells A cells Anti-B Anti-A
O pos pos neg neg
A pos neg neg pos
B neg pos pos neg
AB neg neg pos pos

ABO Discrepancies
 ABO discrepancies happen when there is no match in

results between forward and reverse grouping.
 ABO discrepancies are usually technical in nature and

can be simply resolved by correctly reporting the testing
and carefully checking reagents with meticulous reading
and recording of results.
ABO Blood Grouping Discrepancies
 Missing reactivity?
 apparent absent antigen
 apparent absent antibody
 Additional reactivity?
 apparent extra antigen
 apparent extra antibody

ABO Blood Grouping Discrepancies
Most Common Causes
 Patients
 decreased antibody levels
 physiologic - newborns, elderly
 immunodeficiency - congenital or acquired
 auto- or alloantibodies
 recent transfusion
 Donors
 unusual blood group - genetic polymorphisms
ABO Discrepancies
 There are some ABO discrepancies that can happen due

to technical errors and may lead to false positive or false
negative reactions.
 False positive reactions are due to;

 Un-calibrated centrifuges
 Contaminated reagents
 Dirty tubes or glassware

ABO Discrepancies
 False negative reactions can be due to many causes
 Failure to add serum or reagents

 Use of incorrect reagents or samples
 Cell suspension is too heavy or too light
 Inadequate identification of samples or test tubes

ABO Discrepancies
 Group I discrepancies
These discrepancies are between forward and reverse
grouping due to weak reaction or missing antibodies.
These kind of discrepancies are the most common.
The reason for the missing antibody or weak reaction is

that the patient has depressed antibody production or
cannot produce the ABO antibodies.
ABO Discrepancies
 This type of discrepancy can be seen in new born

infants, elderly patients.
 Patients with lymphoma.
 Patients using immunosuppressive drugs.
 Patients with immunodeficiency disease, BM transplant.

ABO Discrepancies
 Resolving discrepancies
 Eliminate all technical errors

 Enhancing the reaction in reverse grouping
 Incubate the patient’s serum with reagent cells at room
temp. for 15 mins.
ABO Discrepancies
Group II discrepancies
grouping due to weak reaction or missing antigens.
This group is the least one. Can be caused by some
subgroups of A or subgroups of B or both.
Also it can be present in patients with leukaemia and

hodgkin’s disease.
ABO Discrepancies
Group III discrepancies

grouping due to protein or plasma abnormalities.
These can be caused by elevated levels of globulin from

certain diseases such as multiple myloma, hodgekin’s
lymphoma. Some are caused by (Rouleaux formation).
ABO Discrepancies
 Rouleaux or red cells result from a stacking of

erythrocytes that adhere in a coin-link fashion giving the
appearance of agglutination.
 To resolve this kind of problem, washing the patient’s

red cells with saline or adding a drop or two of saline to
the tube in case of rouleaux formation.
 If the agglutination is true red cell clumping will remain.
 Cord blood must be washed 6-8 times in forward

grouping ONLY.
ABO Discrepancies
Group IV discrepancies
 These kind of discrepancies are between forward and
reverse groping due to miscellaneous problems.
 Polyagglutination can occur due to exposure of hidden

erythrocyte Ag. (T antigen) in patients with bacterial or
viral infection.
 Bacterial contamination in vitro or vivo produces an

enzyme that alters and exposes the hidden Ag. on red cell
leading to T activation.
ABO Discrepancies
 Some examples of discrepancies
 Example 1
 Forward grouping: anti-A =O, anti-B =O, anti-AB= O
 Reverse grouping: A1 cells= O, B cells =O
 Blood group:
 O
 Possible discrepancy:
 Missing Ab. Or group I discrepancy
Grouping
Forward Reverse
Missing/Weak Extra Mixed Field Missing/Weak Extra
Young
Cold
A/B Subgroup Acquired B O Transfusion Elderly
Immunocompromised Autoantibody
Disease Bone Marrow Cold

B(A) Phenotype
(cancer) Transplant Alloantibody
Rouleaux Rouleaux
May cause all +
reactions
Anti-A1

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ABO Discrepancies

Basic ABO Grouping

O pos pos neg neg

A pos neg neg pos

B neg pos pos neg

AB neg neg pos pos

 ABO discrepancies happen when there is no match in

 ABO discrepancies are usually technical in nature and

 apparent absent antibody

 apparent extra antibody

 physiologic - newborns, elderly

 immunodeficiency - congenital or acquired

 There are some ABO discrepancies that can happen due

 False positive reactions are due to;

 Dirty tubes or glassware

 False negative reactions can be due to many causes

 Failure to add serum or reagents

 Cell suspension is too heavy or too light

 Inadequate identification of samples or test tubes

These kind of discrepancies are the most common.

The reason for the missing antibody or weak reaction is

 This type of discrepancy can be seen in new born

 Patients with lymphoma.

 Patients using immunosuppressive drugs.

 Patients with immunodeficiency disease, BM transplant.

 Eliminate all technical errors

Also it can be present in patients with leukaemia and

Group III discrepancies

These discrepancies are between forward and reverse

These can be caused by elevated levels of globulin from

 Rouleaux or red cells result from a stacking of

 To resolve this kind of problem, washing the patient’s

 If the agglutination is true red cell clumping will remain.

 Cord blood must be washed 6-8 times in forward

 Polyagglutination can occur due to exposure of hidden

 Bacterial contamination in vitro or vivo produces an

 Some examples of discrepancies

Missing/Weak Extra Mixed Field Missing/Weak Extra

Disease Bone Marrow Cold

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