1 Blood Bank Practical Sheets

Practical NO (9): E- Compatibility Test (Cross-Matching)

1. Objectives:
At the end of this practical you will be familiar to:
1. The intended use and the principle of cross-match.
3. Types of cross-match procedure.
2. Materials and methodology of the test.
4. The intended use of each phase of cross-match procedure.
5. Interpretation of results.
6. Test limitation and precautions.

2. Compatibility Test (Cross-matching):

2.1:Intended use:

Blood bank personnel are responsible for providing

serologically compatible blood within an appropriate time frame. A
cross-match test is performed for patients who need replacement
or compensating 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).

2.2: Principle:

There are two types of cross-matching procedures:

1) 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 or
decreased cell survival of donor cells.
2) 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.

 Testing of the recipient's serum with donor cells (major cross-

match) is required because it is the best available way to detect
antibodies in the recipient's serum that might damage
transfused red cells and cause hemolytic transfusion reactions.

Prepared by: Samah Awad AL-Subhi

2 Blood Bank Practical Sheets

2.3: Procedure of Major cross-match:

2.3.1:Sample:

1) EDTA and a segment of a donor unit to do forward grouping

and cross match.

2) Serum of recipient to do ABO reverse typing, Ab screening test,

and cross match.

2.3.2: Reagents, Equipment, and Supplies:

 Indelible marking pen.

 Glass test tubes.
 Disposal pipettes.
 Wash bottle with physiologic saline.
 AHG reagent (Coomb's serum).
 Test tube rack
 37oC incubator
 22% Bovine albumin or LISS
 Coomb's Control Cells.
 Centrifuge.
 Lighted agglutination viewer.
 Glass slide.
 Microscope.

2.3.3: Method:

1. Verify that patient information on the sample matches

information on the worksheet.

2. Perform forward and reverse typing on recipient's specimen

according to ABO and D lab procedure and Perform Ab screening
test on recipient's specimen according to Ab screening test lab
procedure then record results.

3. Whenever possible the blood selected for cross-match should be

of the same ABO and D group as that of the recipient.

4. Select one segment of donor units, Perform forward typing on

segment of donor units according to ABO and D lab procedure and
record results.

5. Prepare cell suspensions from segment of donor units.

Prepared by: Samah Awad AL-Subhi

3 Blood Bank Practical Sheets

6) Saline Phase (for detection of ABO incompatibility in saline at

room temperature):

a. Add 2-3 drops recipient's serum in all appropriately labeled

tubes and one drop of donor cells.

b. Centrifuge for 15 seconds at 3400 rpm.

c. Gently re-suspend cell button and examine for hemolysis or

agglutination with the aid of the lighted agglutination viewer
(Macroscopically) and microscopically. Then record the reactions
immediately.

7) Albumin or LISS Phase (for detection of antibodies agglutinating

at 37°C in saline):

a. Add 2 drop of LISS in each test tube and incubator for 10

minutes .If LISS not available then add 22% bovine albumin and
increase the incubation period to 30-60 minutes.

b. Centrifuge for 15 seconds at 3400 rpm.

C. Examine for hemolysis or agglutination as usual and record

reactions immediately.

8) Coomb's Phase (for detection of immune IgG antibodies that have

sensitized donor's cells and only agglutinated by using anti-globulin
(AHG) reagent):

a. Wash the tube three times with normal saline.

b. Add two drops of Coombs serum (AHG) to the tube.

c. Centrifuge for 15 seconds at 3400 rpm.

e. Examine for hemolysis or agglutination as usual and record

reactions immediately.

9) Confirm negative result by adding one drop Coombs control cells

(CCC) to tube showing no agglutination and centrifuge for 15 seconds
at 3400 rpm.

10) Gently re-suspend and examine for agglutination. Agglutination

should be present in this step or the test is invalid.

11) Write a check mark in the appropriate place on the worksheet as

the tube has been read.

Prepared by: Samah Awad AL-Subhi

4 Blood Bank Practical Sheets

12) Discard all materials in the appropriate trash containers.

2.3.4:Results Interpretation:

1) If the tube becomes positive at any phase in cross-match

procedure prior to the addition of CCC, the units are
interpreted as “incompatible”, record as “incomp” on your
sheet.

2) If the tube has remained negative throughout the cross-

match procedure, and the check cells is “positive”, the units
are interpreted as “compatible”, record as “comp” on your
sheet.

2.4: 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) In an emergency, blood is issued after completion of the first

stage of a standard cross-match to exclude ABO incompatibility.
This must be indicated on the label attached to the blood bag, all
this emergency cross-match procedure takes about 10 minutes
only.

3) If the antibody screen test is negative, blood is issued after

complete the first stage of a standard cross-match. However, if
antibody screen test is positive, all standard cross-match stages
should be performed.

Prepared by: Samah Awad AL-Subhi

5 Blood Bank Practical Sheets

Worksheet of practical NO (10)

Number of sample:

1- Determine blood group of donor & recipient:

Donor Recipient

Forward typing

__
Reverse typing

2- Antibody screen test: ______________________

3- Antibody identification test if needed: ______________________

4- Selecting suitable Blood Bag: ______________________

5- Antigen typing on donor segment if needed: _________________

6-Results of Compatibility Test (Cross-matching):

RT phase 37 oC phase AHG phase ✔

Cross-
matching tube

Results Interpretation:

Prepared by: Samah Awad AL-Subhi

6 Blood Bank Practical Sheets

Exercise:
Study Questions:

1. What is the primary purpose of the cross-match procedure?

2. Patient need blood transfusion due to massive bleeding, how YOU

can deal with this emergency case?

3. Define “major cross-match”. Why this method is a routine use in

blood bank?

4. The patient specimen must be less than 72 hours old for

compatibility testing, why?

5. Draw a picture organizational flow chart for performing the cross-

match procedure?

Prepared by: Samah Awad AL-Subhi