CHAPTER -SEVEN

The Cross-Match
(compatibility testing)

CH
Content

 Cross matching

 Types of Cross Match

 Steps for compatibility testing

 Choice of Blood for cross-match

 Procedure for cross-match

Learning Objectives

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
7.1 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)
Cross-Matching(#1)

 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 and
 Technical or labeling errors (clerical errors)
Cross-matching(#2)

 Will not:
 prevent immunization of the patient

 guarantee normal survival of transfused

erythrocytes

 detectall unexpected antibodies in a patient’s

serum.
7.2 Types of 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
Types of Cross Match(#1)

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
7.3. Steps for compatibility testing

 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).
Steps for compatibility testing(1)

 Finally, during the actual transfusion :

 careful observation of the recipient’s vital signs and

 posttransfusion hematocrit and Heamoglobin levels

must be considered.
7.4.Selection of Blood for cross-match

 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.
Choice of blood for donation

 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
Choice of blood for donation(1)

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.
Choice of blood for donation (2)

 When cross-matching is carried out, the serum

is tested against the cells.

 The serum should be fresh, that is not more than

48hours old, to make sure that it contains
complement.
Choice of blood for donation(3)

 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..
Choice of blood for donation(4)

 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
1. Saline tube

 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.
2. Albumin tube

 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
 the antibodies react in albumin or any other
protein medium
Albumin tube…

 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.
3. Anti-human globulin tube

 A more concentrated suspension of red cells is

mixed with the patient’s serum and incubated at
370C and then AHG is added.

 A postive test detects the presence of antibodies of:

 rhesus, kell, kidd, S and Lewis

 Anti globulin is essential for detection anti-Duffy

7.5. Procedure for cross-match
7.5.1.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
Cross match (Standard)…

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
Cross match….

Type of specimen

 Serum (plasma) not older than 48 hrs

 Washed cells (20-30% and 2-5%)
Cross match (Standard)…

Equipments and reagents

 Test tubes
 Centrifuge
 Microscope
 Microscopic slide
 Normal saline
 20% albumin
 AHG (Coombs reagents)
Procedure

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 ofdonor’s cells
Cross match (Standard)…

2. Incubate tube 1at 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 globulins are removed from the cells.

4. And make a 3% saline suspension of the washed

cells in a tube.
Cross match (Standard)…

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 heamolysis

macroscopically and microscopically for
agglutination.
Cross match (Standard)…

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.
7.5.2 Emergency cross match

 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.

Cross match (Emergency)…

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
Cross match (Emergency)…

Type of specimen

 Serum(plasma) not older than 48 hrs

 Washed cells (2-5%)
Cross match (Emergency)…

Equipments and reagents

 Test tubes
 Centrifuge
 Microscope
 Microscopic slide
 Normal saline
 20% albumin
Procedure:

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
Cross match (Emergency)…

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.
Cross match (Emergency)…

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.
7.5.3. Rapid direct slide cross match
(Request for un cross matched blood )

 Takes only 3 or 4 minutes

 Plasma is used instead or serum.

 Not safe and must only used in extreme

emergencies

 Standard cross match should be carried out while the

transfusion is in progress.
Procedure

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
Cross match (Rapid)…

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.
Cross match (Rapid)…

Sources of errors in cross-matching

 Rouleaux
 Auto agglutinins
 Infected donor cells
 Over centrifugation
 Dirty glass wares etc..
Review Questions

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.
References

1.Immunohematology for medical laboratory science

students,Yayehyirad T. and Misganaw B., Upgraded
lecture note.2008
2.Basic and applied concepts of Immunohematology,
2nd ed. Kathy D.Blaney and Paula R.Howard,2009
3. Blood banking and transfusion medicine: basic
principles and practice. Christopher D.Hilliyer et al.,
2nd ed.2007.
4.Safe blood donations, Module 1 WHO.2002

41
References…

5.Screening for HIV and other infectious agents,

Module 2, WHO. 2002
6.Blood group serology. Module 3 WHO.2002
7.Guidelines and principles for safe blood transfusion
practice, Introductory module. WHO 2002.
8.Immunohematology: Principles and Practice
Quinley. 2nd ed.1998.
9.AABB Technical Manual .15th Edition.2005

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