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Albano, Astorga, Dumangon, Fregil, Gutierrez,


Pretransfusion Compatibility MINOR Crossmatch

Testing Donors serum+ Patients
red cells (DS-PR)
a.k.a. Compatibility Testing

Series of testing procedures and

processes with the ultimate objective
of ensuring the best possible result
(safety) of a blood transfusion Phases of Crossmatching
The primary objective of the Immediate Saline Spin Phase
crossmatch test is to detect the
presence of antibodies in the Also known as Protein /
recipients serum that could destroy Albumin / Room
transfused RBCs. Temperature phase

Accomplished by mixing the

Crossmatch testing /
patients serum with donors
Crossmatching RBCs (Major) and the donors
serum with patients RBCs
Traditionally meant the testing of the
(Minor) then centrifuge it
patients serum with the donor RBCs
including an antiglobulin phase or
simply an immediate spin phase to Absence of agglutination or
confirm ABO compatibility hemolysis indicates
Two main functions of serologic
crossmatching: Detects:
It serves as a final check of ABO Incompatibility in ABO
compatibility between donor system
and patient.
Incompatibility due to
It may detect the presence of cold antibodies
an antibody in the patients
serum that will react with Prozoning anti-Rh
antigens on the donor RBCs but antibodies are detected
that was not detected in Ab in a serum albumin
screening because the mixture on immediate
corresponding Ag was lacking centrifugation
from the screening cells.
Thermo Phase
It is divided into 2 parts:
Also known as Incubation
MAJOR Crossmatch phase

Patients serum + In this phase, the tubes showing

Donors red cells (PS-DR) no agglutination in the
Albano, Astorga, Dumangon, Fregil, Gutierrez,

immediate spin phase will be Detects

incubated for 30 minutes at
37C water bath (10 mins if LISS Anti-Fya, -Jka, -K
is used instead of 22% BSA)
Antibodies present in
then centrifuge
acquired hemolytic
No agglutination or hemolysis anemia
indicates compatibility
Antibodies in Rh system
Detects which react only in the
AHG test (called 3rd order
Incompatibility due to or cryo-agglutinoid
presence of low titered antibodies)
Causes of Positive Results in
Certain Rh antibodies
(anti-C, E and some D)
Incorrect ABO grouping of the patient
Antihuman Globulin Phase
or donor
Also known as Coombs phase
An alloantibody in the patients serum
For greater sensitivity, AHG reacting with the corresponding
containing both anti-IgG and antigen on donors RBCs
anti-complement may be used
An autoantibody in the patients
blend of rabbit anti-IgG and serum reacting with the corresponding
murine monoclonal anti- antigen on donors RBCs
Prior coating of the donors RBCs with
In this phase, the cells of tubes protein, resulting in positive AHG test
showing no agglutination in the
Abnormalities in the patients serum
previous phase are washed with
NSS thrice. Next, AHG is added Albumin/Globulin (A/G) ratio
and then centrifuged. imbalance
Anti-Human Globulin AntiIgG, Presence of high molecular
-C3d; polyspecific acts as a link weight dextrans or other plasma
between the antibody and/or expanders
complement coating of
neighbouring red blood cells Antibody against additives in
and induces agglutination. the albumin reagents
Uncoated red blood cells will not
agglutinate. Problems Encountered
No agglutination or hemolysis Rouleaux formation
indicates compatibility
Albano, Astorga, Dumangon, Fregil, Gutierrez,

In some diseases
macroglobulinemia Procedures to shorten cross-
matching time
Certain synthetic plasma
expanders like dextran Test should be performed at 37C

Fibrinogen LISS instead of saline

Pan agglutination spontaneous clumping 2% suspension of donor red cell

of cells against a given serum
Enhancing agents (albumin, enzymes)
Bacteriogenic pan agglutination
(Huebener-Thomsen Polycation or polybrine
Friedenreich phenomenon).
Reaction takes place at 20C not
at 37C.

Non-bacteriogenic pan
agglutination caused by
acquired hemolytic anemia or
by rare specific antibodies.
Reaction takes place at 37C.
DAT is always (+).


Cold agglutinins

Whartons jelly


Ag-ab deterioration

Presence of other immediate spin-

reactive antibodies

Hyperimmune ABO antibodies

Procedure is not performed properly

Infants specimens are tested

Contaminants in the test system


reported to eliminate some of
the false-positive reactions
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Protein Phase 0 0 Blood is compatible in both major
1 Thermo Phase 0 0 and minor crossmatch. Blood is safe for
AHG Phase 0 0 transfusion.
Protein Phase + 0 Blood is incompatible in major
Thermo Phase 0 crossmatch but compatible in minor
crossmatch. Blood is not safe for
AHG Phase 0 transfusion
Protein Phase + + Blood is incompatible both in
3 Thermo Phase major and minor crossmatch. Blood is not
AHG Phase safe for transfusion.
Protein Phase 0 + Blood is compatible in major
Thermo Phase 0 crossmatch but incompatible in minor
crossmatch. Blood can be transfused but
AHG Phase 0 with caution.
Table 1.1 Summary of Interpretation of Crossmatching Results