Practical session no 9

D- Compatibility Test (Cross-Match)

Done by nada fallatah,2016

 The cross match is an in vitro procedure to determine
serologic compatibility between the donor red cell and
recipient serum.
 It is performed for patients who need replacement for their
blood volume due to anemia or active, massive bleeding.
 The purpose of cross matching is to prevent the transfusion of
incompatible cells (post-transfusion reaction).
Selection of Packed Red Blood Cells (PRBC) and platelet for
transfusions:
When a select blood group of recipient should be compatible
with blood group of donor. When the blood of the recipient's
ABO group is unavailable, transfusion with an alternate group,
as shown, is acceptable.
 A
O AB
B

Figure 1: Selection of Packed Red Blood Cells (PRBC) and platelet for
transfusions flow chart.

Universal donors for packed red blood cells:-

It is blood group O with Rh negative donors. It lacks all ABO
and D antigens.
The universal recipient for packed red cells:-
It is blood group AB with Rh positive. It lacks of ABO
antibodies they will be compatible with all red blood cell types.
Selection of Plasma for transfusions: The rules are
inverted !

O AB
B

Figure 2: Selection of Plasma for transfusions flow chart

Universal donors for plasma transfusions:
It is blood group AB due to lack all ABO antibodies.
Universal recipient for plasma transfusions:
It is blood group O which has all ABO antibodies present
All plasma products will be compatible.
 Intrauterine transfusion:
Blood transfusion for intrauterine should be compatible with
the maternal blood group.

Blood transfusion of new-born:

 If ABO/Rh of the fetus has been in umbilical blood sample,
then the blood group–specific could be given.
 If ABO/Rh of the fetus are not known should be given O
negative.
 Blood transfusion should be as fresh and no older than 7
days.
 Compatibility testing using the mother's serum sample.
 Compatibility testing in emergencies:-
1. Blood is issued after completion of the first stage
of a standard cross-match to exclude ABO
incompatibility.
2. This is emergency cross-match procedure takes
about 10 minutes only.
3. If not time, should give blood group O negative.
 2: Principle:
Cross match become part a series of pre–transfusion test
includes: ABO/Rh grouping, antibody screening for both
donor and recipient.
There are two types of cross-matching procedures:

 The major cross-match involves testing the recipient's

serum with donor cells to determine whether the recipient
has an antibody which may cause a hemolytic transfusion
reaction.
 The minor cross-match involves testing the recipient's
cells with donor plasma to determine whether there is an
antibody in the donor's plasma directed against an antigen
on the recipient's cells.
3: Sample:
1) EDTA or a segment of a donor unit.
2) Serum of recipient.
4: Reagents, Equipment, and Supplies:
 Glass test tubes.

 Disposal pipettes.

 Wash bottle with physiologic saline.

 AHG reagent (Coomb's serum).

 37oC incubator

 22% Bovine albumin or LISS

 Coomb's Control Cells.

 Centrifuge.
5: Method:
1- Perform ABO/Rh type (forward and reverse) and antibody
screening on the recipient's specimen.
2- Whenever possible the blood selected for cross-match
should be of the same ABO and D group as that of the
recipient.
3-Select one segment of donor units, Perform ABO/Rh
forward typing on a segment of donor units.
4- Prepare cell suspensions from a segment of donor units
Saline Phase (for detection of ABO incompatibility in
saline at room temperature):
5- Add 2-3 drops recipient's serum in all appropriately
labeled tubes and one drop of donor cells.
6- Centrifuge for 15 seconds at 3400 RPM.
7- examine for hemolysis or agglutination and record the
result.
Albumin or LISS Phase (for detection of antibodies
agglutinating at 37°C in saline):
8- Add 2 drops of 22% bovine albumin and incubation for to
30 minutes.
9-Centrifuge for 15 seconds at 3400 RPM.
10-Examine for hemolysis or agglutination as usual and record
reactions immediately.
Coomb's Phase (for detection of immune IgG antibodies):
11- Wash the tube three times with normal saline
12- Add two drops of Coombs serum (AHG) to the tube.
13-Centrifuge for 15 seconds at 3400 RPM.
14- Examine for hemolysis or agglutination as usual and
record reactions immediately.
15-Confirm negative result by adding one drop Coombs
control cells (CCC) to tube showing no agglutination and
centrifuge for 15 seconds at 3400 RPM.
16- Gently re-suspend and examine for agglutination.
Agglutination should be present in this step or the test is
invalid.
6:Results Interpretation:
1) If the tube becomes positive at any phase in the cross -
match procedure, the units are interpreted as
“incompatible”.
2) If the tube has remained negative throughout the cross-
match procedure, and the check cells are “positive”, the
units are interpreted as “compatible”.

Interpretation

Agglutination No agglutination
(incompatible) (compatible)
 7: Test limitation and Precautions:
1) The patient specimen must be less than 72 hours
old for compatibility testing so that it represents
the current immunologic status of the patient.

2) If the antibody screen test is negative, blood is

issued after completing the first stage of a
standard cross-match which is called “Immediate
Spin Cross-Match”. However, if the antibody
screen test is positive, all standard cross-match
stages “Extend Cross-Match” should be
performed.
8: Adverse effect of blood transfusion reactions

Is it the destruction of RBC s inside human body due to blood

transfusion which is happening immediately or period of
time due to an immune or non-immune reactions.
Clinical consequence: It is very variable!!!
Ineffective transfusion

hemodynamic Shock with intravascular haemolysis
1-There are many ways in which a recipient can be
adversely affected by a blood transfusion:

 Bacterial or Viral or Parasitic infection.

 Non-hemolytic febrile reaction (NHFTR).
 Allergic reaction.
 Post-transfusion purpura (PTP).
 Transfusion-associated acute lung injury (TRALI).
 Transfusion-associated graft versus host disease (TA-GvH)
 2- laboratory investigation:-
1. Check that the patient’s details are identical on both the
pre- and post-transfusion samples, and on each request
form to compare current ABO and Rh.
2. Reaper the group on pre & post transfusion sample and on
the donor blood and repeat the cross match.
3. Perform DAT and antibody identification on post
transfusion sample.
4. Check the plasma for hemoglobinemia
5. Perform test for coagulation profile
6. Examine donor sample for bacterial contamination.
7. Examine post transfusion sample of urine for
hemoglobinuria
8. A fourth sample of blood is taken 6h and or 24 h after
transfusion of blood for CBC, bilirubin, methaemalbumin.
3-Management of transfusion reaction:-
1-Hydrocortisone 100mg and antihistamine which alleviate
shock.

2-If sever shock give adrenaline 1:1000.