 Cross matching

 Types of Cross Match

 Steps for compatibility testing

 Choice of Blood for cross-match

 Procedure for cross-match

At the end of this chapter, the student will be able
to:
 Explain the cross-match and its primary purpose.

 Describe the constituents of the major and

minor cross match.
 Select appropriate blood for cross-match.

 List the types of antibodies that can be

encountered at various phases of a cross-match.
 Perform cross matching
 It is a test to determine the compatibility
between recipient’s blood and donor’s blood.
 It is a procedure performed before transfusion
to select donor’s blood that will not cause any
adverse reaction, (hemolysis /agglutination)
 The test is important to see whether there is
danger or not if the donor’s blood is transfused
to the recipient.
 Helps the patient to receive maximum benefit from
the transfusion of red cells
 Main purpose is to prevent a transfusion
reaction that may result due to:
 Unexpected antibodies in the patient’s /donor’s
serum
 Some ABO incompatibilities
 Technical or labeling errors (clerical errors)

 Increase in vivo survival of red cells

 Double checks for ABO errors
 Another method of detecting antibodies
 Transfusion reaction – is the term used to
describe the clinical effects
caused by the destruction of red cells of
recipient by the donor’s antibodies or a
donor’s red cell’s by the recipient
antibodies during an incompatible
transfusion.
But :
 Will not prevent immunization of the patient

 Will not guarantee normal survival of transfused

erythrocytes

 Will not detect all unexpected antibodies in a

patient’s serum.

 Will not detect all recipient antibodies to donor

antigens
Major and Minor cross-match
Major cross-match:
 Involves mixing recipient’s serum with the
donor’s red cells.
 Is much more critical for assuring safe
transfusion than the minor compatibility test.
 Called major b/c the Abs in the recipient’s serum
are most likely to destroy the donor’s RBC
Minor cross match:
 Involves mixing the donor’s serum with
patient’s red cells
 Called minor because

 Any Ab in the donor’s serum will be diluted by the

large volume of the recipient’s blood

 The destructed RBCs of the patient may be

compensated by the transfused RBC of the donors
 Accurate Patient Identification

 Proper sample collection and handling

 Review of the recipient’s past blood bank

records
 Careful ABO/Rh determination

 Antibody screening of the recipient (cross

matching of the donor unit).
 In cases when the recipient possess a clinically
significant Antibody, donor units must be:
 Screened for the corresponding Ag and should be
negative
 Cross- matched

 Finally, during the actual transfusion :

 Careful observation of the recipient’s vital signs and

 Post transfusion hematocrit and Hemoglobin levels

must be considered.
 The blood selected for cross-match should be of
the same ABO and Rh (D) group as that of the
recipient.
 However, Rh positive recipients may receive
either Rh positive or Rh negative blood.
 Whenever possible blood of the patients own blood group should
be given.
 Otherwise the following rules should be applied.
Group A patient.
- Should receive group A blood, if not available group O
Group B patient.
- Should receive group B blood, if not available group O
 Group O patient.
 Can only receive group O blood
Group AB patient.
 Should receive from group AB, if not possible can receive
blood from group A,B, and O.

 Rh-pos blood should not be given to Rh-neg women

of childbearing age.
 Transfusion of Rh-neg male patients and female
patients beyond menopause with Rh-pos blood is
acceptable as long as no preformed anti-D is
demonstrable in the sera.
 When cross-matching is carried out, the
serum is tested against the cells.

 The serum should be fresh, that is not more

than 48 hours old, to make sure that it
contains complement.
 When deciding on methods for cross-
matching, the following conditions are
required for Ag-Ab reactions.

 The right Temperature.

 Suitable surrounding medium
 Antigen-Antibody ratio etc..
 The safe cross-matching of blood requires
that the donor’s cells be mixed with the
patient’s serum in three separate tubes, using
:

1. Saline
2. Albumin
3. Anti-human globulin reagents
 The red cells from the donor are suspended in
saline and mixed with the patient’s serum.
 Show the presence of any complete
antibodies
 Agglutination in the saline tube is usually
caused by:
 anti-A or anti-B antibodies and
 Occasionally by Lewis, MNSs, Lutheran and kell
antibodies.
Remember that
5% means

SALINE added
IgM antibody
 The red cells from the donor’s suspended in saline, are mixed
with the patient’s serum, and albumin is added.
 The tube is incubated at 370C
 Shows the presence of any incomplete antibodies
 Agglutination in the albumin tube is often caused by:
 The Rhesus antibodies,
 Lewis, MNSs, Lutheran and P antibodies, and
 Occasionally by anti-kell.

 Reaction caused by anti- A or anti- B antibodies usually occur

in albumin as well as in saline.
 A more concentrated suspension of red cells
is mixed with the patient’s serum and
incubated at 370C and then AHG is added.
 A positive test detects the presence of many
antibodies of:
 Rhesus, Kell, Kidd, S and Lewis

 Anti globulin is essential for detection anti-

Duffy
 AHG is for
Incomplete or
IgG
antibodies
Standard cross-match
 Is cross-match that is performed in three
tubes (Saline, albumin and AHG) within 45 to
60 minutes

Clinical significance
 Detects unexpected (irregular) antibodies in
the recipient/ donor serum
Principle

 Serum of the recipient / donor is tested

against the red cells of the donor/ recipient
under different conditions in order to
establish their compatibility
Saline phase will include

Saline at 22oC for IgM

Saline + Albumin at 37oC

for IgG
Type of specimen

 Serum (plasma) not older than 48 hrs

 Washed cells (20-30% and 2-5%)
Equipments and reagents
 Test tubes
 Centrifuge
 Microscope
 Microscopic slide
 Normal saline
 20% albumin
 AHG (Coombs reagents)
1. Take 3 small tubes mark them 1,2 and 3, and
add to each the following
Tube 1 1volume of patient’s serum
1volume of 3-5% donor’s red cells
Tube 2 1 volume of 20% bovine albumin
1volume of patient’s serum
1volume of 3-5% donor’s red cells
Tube 3 3 volume of patent’s serum
1volume of 20-30% suspension of
donor’s cells
2. Incubate tube 1 at RT for 30 min and Tube 2
for 30 minutes at 370C. Incubate tube 3 for 15
minutes at 370C.
3. After incubation, remove tube 3 and wash
the cells three times with clean saline to
make sure that all the non-bouned globulins
are removed from the cells.
4. And make a 3% saline suspension of the
washed cells in a tube.
5. To one volume of red cell deposit add 2
volumes of fresh diluted antiglobulin
(Coombs) Reagent.

6. Remove tube 1 and 2 and centrifuge with

tube 3 for one minute at 1000 rpm

7. Examine the tube for hemolysis

macroscopically and microscopically for
agglutination.
Results
 No hemolysis or agglutination is seen in tube 1, 2
or 3
 The blood is compatible and can be issued with the
completed cross-match label.

 If there is agglutination or hemolysis in any of

the tubes
 The blood is incompatible, and must not be issued for
the patient.
ABO incompatibility (anti A and Anti-B.)
 Saline tube ………………….Shows strong agglutination
 Albumin tube ………………. Shows agglutination
 Anti- globulin tube ………… Show no agglutination

Rhesus incompatibility (anti-D ,c, e)

 Saline tube …………………….Does not usually show
agglutination
 Albumin tube …………………. Shows agglutination
 Anti-globulin tube ……………. Shows agglutination
Anti- Duffy and anti- kidd incompatibilities
 Saline tube …… Does not usually show agglutination
 Albumin tube … Does not usually show agglutination
 Anti- globulin tube ……………. Shows agglutination

Anti- Lewis incompatibility

 Saline tube …………………….. Shows agglutination
 Albumin tube ………………….. Shows agglutination
 Anti- globulin tube ……………. Shows agglutination
 Performed when there is no enough time to
perform the standard cross match
 Takes about 25 to 30 minutes and

 Does not include antiglobulin test.

Principle
 Serum of the recipient / donor is tested against
the red cells of the donor/ recipient in saline and
albumin medium in order to establish their
compatibility
Type of specimen
 Serum (plasma) not older than 48 hrs
 Washed cells (2-5%)
Equipment and reagents
 Test tubes
 Centrifuge
 Microscope
 Microscopic slide
 Normal saline
 20% albumin
1. Take 2 small tubes, mark them 1 and 2 and
add to each the following

Tube 1 1volume of patient’s serum

1volume of 3-5% donor’s red cells
Tube 2 1volume of patient’s serum
1volume of 3-5% donor’s red cells.
1volume of 20% bovine albumin
2. Leave tube 1 at room temp for 15 minutes
incubate tube 2 for 15 minutes at 370C.

3. Centrifuge both tubes for one minute at 1000

rpm/min

4. Examine the tubes macroscopically for

hemolysis and microscopically for
agglutination.
Results
 If no hemolysis or agglutination is seen in
either tube 1 or 2
 the blood is compatible and can be issued with the
emergency cross match.

 If agglutination or hemolysis is seen in either

of the tubes
 the blood is incompatible and must not be issued
for the patient.
 Takes only 3 or 4 minutes

 Plasma is used instead of serum.

 Not safe and must only used in extreme

emergencies

 Standard cross match should be carried out

while the transfusion is in progress.
1. Take 2 volume of patient’s plasma on a slide

2. Add 1 volume of donor’s whole blood of the

same group as the patient and mix.

3. Leave for 2 minutes and examine

microscopically for agglutination
Results

 If the cells show agglutination

 the blood must not be given and will usually
indicate that the wrong ABO group blood is
being cross marched.
Sources of errors in cross-matching
 Rouleaux
 Auto agglutinins
 Infected donor cells
 Anti- A1
 Over centrifugation
 Dirty glass wares etc..
1. What is cross-matching?

2. What is the purpose of cross-matching?

3. List the types of cross-match with their

constituents

4. List the stages of cross-match and their

respective importance in antibody detection.