ABO DISCREPANCIES

 Occurs when red cell testing does not agree with the expected
serum testing.
 It is suspected when:
o Agglutination strengths of the typing reactions are
weaker than expected
o Expected reactions in ABO red cell testing and serum
testing are missing
o Extra reactions are noted in either the ABO red cell or
serum tests
 It occurs when unexpected reactions occur in forward and reverse
typing. It can be divided into four categories: group I, group II,
group III and group IV.
Group I Discrepancies Resolution:
 Obtaining the patient’s history may resolve this type of
discrepancy, such as a newborn sample that would not have ABO
antibodies in the serum until the child was 4 to 6 months of age.
 Enhance the weak or missing reaction in the serum by incubating
the patient serum with reagent A1 and B cells at room temperature
for approximately 15 to 30 minutes.
 No reaction after centrifugation – incubate at 4C for 15 to 30
minutes
 Auto-control and O cell control must always be tested concurrently
with the reverse typing.

Group II Discrepancies
It is associated with unexpected reactions in the forward grouping due to
weakly reacting or missing antigens. It is the least frequently encountered
discrepancy.

Some of the causes of discrepancies in this group include:

 Subgroups of A (or B) may be present
 Leukemias may yield weakened A or B antigens, and Hodgkin’s
disease has been reported in some cases to mimic the depression of
antigens found in leukemia.
 The “acquired B” phenomenon will show weak reactions with anti-
B antisera and is most often associated with diseases of the
Group I Discrepancies: digestive tract (e.g., cancer of the colon).
 It occurs when unexpected reactions occur in forward and reverse
typing. It can be divided into four categories: group I, group II,
group III and group IV.
 It is associated with unexpected reactions in reverse typing due to
weakly reacting or missing antibodies.
 The patient has depressed antibody production or cannot produce
the ABO antibodies.
 It is a common discrepancy.

Common populations with discrepancies in this group are:

 Newborns (the production of ABO antibodies is not detectable
until 4 to 6 months of age)
 Elderly patients (the production of ABO antibodies is depressed)
 Patients with a leukemia (e.g., chronic lymphocytic leukemia) or
lymphoma (e.g., malignant lymphoma) demonstrating
hypogammaglobulinemia.
 Patients using immunosuppressive drugs that yield
hypogammaglobulinemia.
 Patients with congenital acquired agammaglobulinemia or
immunodeficiency diseases
 Patients with bone marrow or stem cell transplantations (patients
develop hypogammaglobulinemia from therapy and start
producing a different RBC population from that of the transplanted
bone marrow)
 Patients whose existing ABO antibodies may have been diluted by
plasma transfusion or exchange transfusion
 ABO subgroups

Group II Discrepancies Resolution:

  Enhance by incubating the test mixture at room temperature for up
to 30 minutes to increase the association of antibody with antigen
 If negative, incubate the test mixture at 4C for 15 to 30 minutes.
Group III Discrepancies
A type of discrepancies between forward and reverse groupings caused by
protein or plasma abnormalities and result in rouleaux formation or
pseudoagglutination attributable to:
 Elevated level of globulin (Multiple myeloma, Waldenstrom’s
macroglobulinemia, Plasma dyscrasia)
 Elevated level of fibrinogen
 Plasma expanders such as dextran or polyvinylpyrrolidone
 Wharton’s jelly in cord blood
 Unexpected alloantibodies in the patient’s other than ABO
isoagglutinins.
 Panel could be performed with the patient’s serum.
 Once unexpected alloantibodies are identified, A1 and B cells
negative for the corresponding antigen can be used in the reverse
typing or once again the reverse typing can be repeated at 37C if
the ABO isoagglutinins react at this temperature and there is no
interference from the unexpected alloantibody .

Group III Discrepancies Resolution:

 Perform saline dilution or saline replacement technique
 Wash cord cells six to eight times with NSS

Group IV Discrepancies
These discrepancies between forward and reverse groupings are due to
miscellaneous problems and have the following causes:
 Cold reactive autoantibodies in which RBCs are so heavily coated
with antibody that they spontaneously agglutinate, independent of
the specificity of the reagent antibody .
 Patient has circulating RBCs of more than one ABO group due to
RBC transfusion or marrow/stem cell transplant
 Unexpected ABO isoagglutinins
 Unexpected non-ABO alloantibodies

Group IV Discrepancies Resolution:

Potent cold autoantibodies can cause spontaneous agglutination of the
patient’s cells:
 RBCs could be incubated at 37C for a short period, then washed
with NSS and retyped.
 If not successful in resolving forward type, treat patient’s RBC
with 0.01M dithiothreitol to disperse IgM related agglutination
 As for the serum, the reagent RBCs and serum can be warmed to
37oC, then mixed, retested and read at 37C.
 If the reverse typing is negative autoabsorption is done to remove
autoantibody.
Unexpected ABO isoagglutinins in the patient’s serum react at room
temperature – A2 and A2B individuals who can produce naturally occuring
anti-A1 and A1B who produce naturally occurring anti-H
 Specificity of the antibody can be determined by examining the
pattern of reactivity