COMPATIBILITY

TESTING
-Sarah Jane E. Bato
BLOOD TRANSFUSION
PRACTICES
Pre-transfusion
Transfusion
Post-transsfusion
PRE-TRANSFUSION TESTING
Also known as Compatibility testing
The purpose of compatibility testing is to select
blood components that will not cause harm to the
recipients and will have acceptable survival when
transfused.
If properly performed, compatibility tests will
confirm ABO compatibility between the component
and the recipient and will detect the most clinically
significant unexpected antibodies.

The following are important to
assure safe transfusion therapy:
1. Identification of patient and donor and collection
of appropriate samples.
2. Testing of donor sample.
3. Testing of patient sample and review of past
blood bank records.
4. Selection of appropriate donor units.
5. Cross matching
6. Re-identification of the patient prior to infusion of
blood.

COMPONENTS OF
COMPATIBILITY TESTING
Pre-analytical Procedures

Serologic testing

Post-analytical procedures
PRE-ANALYTICAL
PROCEDURES
Positive Patient Identification

Sample Collection (both donors and recipients)

Review of patient history

PATIENT IDENTIFICATION
MISIDENTIFICATION
Most common clerical error.

Errors usually occur when:
1. Blood sample was drawn.
2. Mix-up of samples during handling in laboratory.
3. When transfusion was given.
THINGS TO CONSIDER FOR
POSITIVE PATIENT IDENTIFICATION
1. Blood request form

printing must be legible.
Computerized printouts.


2. Patient wristband identification

Must always be compared with requisition form.
3. Temporary tie tags, wrist band or ankle band.

4. Ask the patients full name and spell it out.
The phlebotomist should never
offer a name and ask to confirm
that it is correct.
SAMPLE COLLECTION
1. PATIENT SAMPLES

2. DONOR SAMPLES
PATIENT SAMPLES
1. SERUM
Serum is more preferable than plasma because
the latter may inactivate complement and also
cause small fibrin clots which may be difficult to
distinguish from true agglutination.

About 10 ml of blood is usually sufficient for all
testing procedures if there are no serologic
problems.
Federal regulations require that fresh serum
obtained from samples less than 72 hours after
collection must be used for antibody screening and
crossmatch testing, if the patient has been
pregnant of transfused within the last 3 months.

2. RED BLOOD CELLS
can be obtained from either clotted or anti-
coagulated samples.
Can be washed to remove plasma or serum which
may interfere with some testing procedure.
A 2-5% Red cell suspension is used for most
serologic tests.

DONOR SAMPLES
1. RED BLOOD CELLS
Donor cells can be obtained from the segment in a
number of ways that permit several procedures to
be performed from the same segment.

Use a lancet to make a tiny hole through which a
single drop of blood can be expressed easily.
Another technique is to cut the segment next to the
cell pack and use an applicator stick to remove
cells, or express a drop by squeezing the tubing.
The segment should be stored with the cut end
down in a test tube to minimize contamination but
never be emptied in it.
REVIEW OF PATIENT
HISTORY
Look at recipients records for any prior
unexpected antibodies.

Previous transfusion reactions.

SEROLOGIC TESTING
ABO and Rh typing

Antibody screening

Cross matching
ABO and Rh TYPING
In the ABO typing, the forward and reverse MUST
match
FORWARD TYPING

REVERSE TYPING
In the Rh typing, the control must be negative

Both of these will indicate what type of blood should be given

ANTIBODY SCREENING
The antibody screen will detect the presence of
any unexpected antibodies in patient serum

ANTIBODIES REGARDED AS ALWAYS BEING
POTENTIALLY CLINICALLY SIGNIFICANT
ABO
Rh
Kell
Duffy
Kidd
SsU
ANTIBODIES THAT MAY SOMETIMES BE CLINICALLY
SIGNIFICANT
Lea
MN
P1
Lutheran
Cartwright
ANTIBODIES THAT RARELY, IF EVER, ARE CLINICALLY
SIGNIFICANT
Leb
Chido/Rodgers
York
Sda
Xga
Bg
HTLA
CROSSMATCHING
TWO MAIN FUNCTIONS OF CROSSMATCHING:

1. It is the final check of ABO compatibility between
donor and recipient.

2. It may detect the presence of an antibody in the
patients serum that will react with antigens or the
donor red blood cells but that was not detected in
antibody screening because the corresponding
antigen was lacking from the screening cells.
2 TYPES OF CROSSMATCH
TESTS
1. Major routinely performed in labs
Mixing of the patients serum with the donor red
blood cells.

2. Minor not required by AABB since 1976
Mixing of donors plasma with patients red
blood cells.

MAJOR CROSSMATCH
Donor RBCs
(washed)
Patient serum
No agglutination ~ compatible
Agglutination ~ incompatible
WHOLE BLOOD
WITH SEGMENT
ATTACHED
For the major test, a 2 to 5% red cell suspension in
saline should be prepared from a sample taken
from an integral segment on the unit selected.

The cells may be washed at least one time to
eliminate plasma and anticoagulant, which may
cause interference in the results.

The ratio of patients serum to donors red blood
cells should be at least 2:1, or equivalent to 2
drops od serum to 1 drop of a 3% red cell
suspension dispensed from droppers of equal size.
SALINE TECHNIQUE
1. Label 1 tube for each donor sample to be tested.
2. Put 2 drop of patients serum in labeled tube.
3. Add 1 drop of 2-5% saline suspended red cells
of donor
4. Mix and incubate for 5-10 min. (spin method) or
incubate for 30-60 min (sedimentation method)
at RT.
5. Centrifuge at 1000 rpm for 1 min. in spin method
(after 5-10 min. incubation);centrifugation is
optional in sedimentation method.

6. Read the result, observe for hemolysis and
agglutination.
7. Negative result should be confirmed under
microscope.
Interpretation
Agglutination or hemolysis indicates a positive result
(incompatible)

ANTI-HUMAN GLOBULIN TEST
(IAT)
1. Put 2 drops of patients serum in a labeled tube.
2. Add 1 drop of 2-5 % saline suspended red cells
of donor.
3. Incubate for 30-60 min at 37 C
4. Centrifuge at 1000 rpm for 1 min, check for
hemolysis/agglutination
5. If there is no hemolysis/agglutination, wash the
cells three times with normal saline.

6. Perform IAT test
Add 2 drops of polyspecific AHG serum to
washed cells
Centrifuge at 1000 rpm for 1 minute
See for agglutination
7. Add IgG coated red cells to negative AHG test.
8. Centrifuge and check for agglutination - if there
is no agglutination test is invalid.
INTERPRETATION
Hemolysis or agglutination at any stage indicates
incompatibility.


METHODS USED TO ENHANCE
ANTIGEN-ANTIBODY
REACTIONS
ALBUMIN

LOW IONIC STRENGHT SOLUTION (LISS)

POLYBRENE (P-AHG)
CAUSES OF POSITIVE RESULTS
IN MAJOR CROSSMATCH
1. Incorrect ABO grouping of the patient of donor
2. Alloantibody in the patients serum reacting with
the corresponding antigen on donor red cells.
3. Autoantibody in the patients serum reacting with
the corresponding antigen on donor red cells.
4. Prior coating of the donor red cells with protein,
resulting in a positive AHG test.
5. Abnormalities in patients serum.
6. Contaminants in the test system.
CROSSMATCHES WILL:
Verify donor cell ABO compatibility
Detect most antibodies against donor cells

CROSSMATCHES WILL NOT:
Guarantee normal survival of RBCs
Prevent patient from developing an antibody
Detect all antibodies
Prevent delayed transfusion reactions
Detect ABO/Rh errors