Crossmatching

:Major and Minor Cross

matching
Caya, Jay G.
Puntukan, Nurdeza N.
BSMT-4A

Objectives:
To understand the principle of cross match
procedure and significance of compatibility
tests.

Reference:
immediate_spin_crossmatch by
marilyncollins (PDF file)
Practical Blood banking by Dr. Marwan
Ibrahim
6. Crossmatch_Carolyn Ragland (PDF file)
clin1XM_Terry Kotrla (PDF file)
COMPATIBILITY_TESTING_7_8 (ppt)
Crossmatching(ppt)

Table of contents:
1.
2.
3.
4.

Principle of Crossmatching
Compatibility Testing
Major and Minor Crossmatching
Phases of Crossmatching:

Immediate Spin phase (IS Phase) Saline Phase

LISS (Low Ionic Strength Solution) or 22% Bovin
Albumin Phase
Anti Human Globulin Phase (AHG/ Coombs
Phase)

I- Principle of crossmatching:
A compatibility test or crossmatch is a
lab procedure to determine before
transfusion IF there is serologic
compatibility between a blood donor
and an intended recipient.

 The patient specimen must be less than

72 hours old for compatibility testing so
that it represents the current
immunologic status of the patient.
Negative results are taken to indicate
compatibility

Purpose of Crossmatching:
Is to PREVENT the TRANSFUSION of
INCOMPATIBLE CELLS.
The transfused red cells will have an
ACCEPTABLE survival rate, and there
will be no significant destruction of the
recipients own red cells.

Compatibility testing
Collecting Patient
Samples
Hemolyzed samples
can not be used for
testing because
hemolysis caused by
activation of
COMPLEMENT

 SERUM or PLASMA may be used for pretransfusion testing.

Most blood bank technologist PREFER
SERUM because PLASMA may cause
small fibrin clots to form which may difficult
to distinguish from true agglutination.

Major and Minor

crossmatching

Major crossmatching
RECIPIENT SERUM is tested
against donor packed cells to
determine if the recipient has
preformed antibodies AGAINST
any antigens on the DONORS
CELLS. This is the required crossmatch prior to release of a unit of
packed cells.

Minor crossmatching
RECIPIENT RED CELLS are tested
AGAINST DONOR SERUM to detect
donor antibodies directed against a
patient's antigens. This is no longer
required. It is assumed that the small
amount of donor serum and
antibodies left in a unit of packed cells
will be diluted in a recipient.

Major crossmatching
The major crossmatch involves testing
the patient's serum with donor cells to
determine whether the patient has an
antibody which may cause a hemolytic
transfusion reaction or decreased cell
survival of donor cells.

Phases
of
Major Crossmatching

1. Immediate Spin Phase (Saline Phase)

2. LISS (Low Ionic Salt solution) 22%
Bovine Albumin Phase
3. AHG Phase or (Coombs Phase)

Note: In blood bank we put step 2-3 in one

step and for agglutination once.

I- Immediate Spin Phase

(Coombs Phase)

I- Immediate Spin Phase

1st wash the donors cell with 0.9 %
NSS and make a 3-5 % cell
suspension.

*WHY???*
Washing
- To help remove unwanted antibodies or antigens
which may interfere with agglutination reaction.

Saline solution / 0.9 % NaCl

- To maintain osmotic balance and prevent the
RBC from shrinking (if NaCl is less than 0.9%),
or swelling up or get burst, which will interfere
with agglutination reaction.

WHY???*
3-5 % cell suspension
- Make the Ab/Ag reaction more clear,
and to avoid false ve and false +ve
results.

Procedure:
1. Add the following to each labelled crossmatch
tube:
-2 drops of patients plasma or serum
-1 drop of the donor cell suspension
2. Mix the contents of each tube.
3. Centrifuge tubes for 15 secs 3000 rpm.
4. Examine for hemolysis. Record if present.
5. Immediately resuspend the cells by gentle
agitation; examine the tubes macroscopically
for agglutination.
6. Grade and record results.

No agglutination =
compatible

Results:
Any DEGREE OF AGGLUTINATION OR
HEMOLYSIS in the crossmatch tubes
during ANY phase indicates
INCOMPATIBILITY and the blood should
not be given.
If AGGLUTINATION OR HEMOLYSIS
occurs in the screen cells tubes, this
indicates the presence of an atypical
antibody and further testing must be done.

II- LISS or Bovine Albumin

Phase

WHY??
solutions such as albumin, polymerized
albumin or low ionic salt solutions
(LISS) are frequently employed to:
increase the sensitivity of the crossmatch
or to reduce the incubation time.
By lowering the ionic strength or
increasing the dielectric constant of the
test medium, these solutions increase
antibody uptake and enhance the strength
of the antigen-antibody reaction.

II- LISS or Bovine Albumin Phase

Procedure:
1. In the same tube just add 2 drops of
albumin, /polymerized albumin or low
ionic salt solutions (LISS).
2. Incubate @ 37C for 20 mins.
3. Wash atleast 2 times.

III- AHG PHASE

*WHY???*
To detect incomplete antibody
like Ig G.

III- AHG PHASE

Procedure:
1. After incubating add 2 drops of AHG
reagent.
2. Centrifuge for 15 secs.
3. Record the result

Checking the crossmatch if it

is valid using check cells.

Procedure
Just add 1 drop of check cells to the same
tube.
Centrifuge for 15 secs.
Record the result.
Note: atleast 2+ agglutination must be
observed to the crossmatch was valid.

For you to understand clearly

lets watch a video

Evaluation:
1. What is the Principle of crossmatching?
2. What is the purpose of detecting Ig G in
AHG phase?
3. What is the primary purpose of
crossmatch procedure?
4. What is the maximum amount of time a
specimen may be used for
pretransfusion testing?

Evaluation:
How is the interpretation of results in
crossmatching?