Name:

Section and Cluster:

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Laboratory Activity No. 8
MLSIMHE23
BROAD SPECTRUM COMPATIBILITY TESTING

I. Introduction
Compatibility testing is a series of procedures designed to ensure the safety of blood
for transfusion. Included in the compatibility test is crossmatching, which is performed prior
to transfusion of blood components containing red cells. A crossmatch testing is divided into
two parts: MAJOR CROSSMATCH, consisting of mixing the patient’s serum with donor red
cells (PS-DR) and the MINOR CROSSMATCH, consisting of mixing the donor’s serum with
patient red cells (PR-DS).

Objectives
At the end of this experiment, the student should be able to:
1. perform major and minor crossmatch with great accuracy;
2. differentiate a compatible from an incompatible blood transfusion reaction;
3. interpret the results of major and minor crossmatch; and
4. learn the significance of doing the procedure.

II. Materials
Apparatus Glassware and other equipment

Wasserman test tubes Marking pen
Pasteur pipettes Gum label
Nescofilm Centrifuge
Gloves

Reagents/Samples

0.85 – 0.9% Normal Saline Solution (in a wash bottle)
2 – 5% Donor RBC suspension
2 – 5% Patient RBC suspension
Patient’s serum
Donor’s serum
Antihuman globulin (AHG) sera
22% Bovine serum albumin (22% BSA)

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III. Procedures

A. Protein/ Albumin/Room Temperature Phase (Immediate Spin)
1. Label 4 wasserman test tubes as M, MA, N, NA.
2. Follow the table below.
CONTENT M MA N NA
Patient’s serum 2 drops 2 drops - -
Donor’s serum - - 2 drops 2 drops
2 – 5% donor RBC suspension 1 drop 1 drop - -
2 – 5% patient RBC suspension - - 1 drop 1 drop
22% Bovine serum albumin - 2 drops - 2 drops

Legend:
M = major crossmatch (PS – DR)
MA = major crossmatch (PS – DR with albumin)
N = minor crossmatch (PR – DS)
NA = minor crossmatch (PR – DS with albumin)

3. Mix the contents and cover with Nescofilm.
4. Centrifuge for 15 seconds at 3,400 rpm.
5. Gently dislodge the cell button and observe for hemagglutination or hemolysis.
6. If no agglutination or hemolysis is observed, only MA and NA tubes will proceed to the
THERMO PHASE.
Note: LISS may be used instead of 22 % BSA if the tubes proceeding to the next step
will be incubated for 10 minutes.

B. Thermo/Incubation Phase (37°C)
1. Incubate MA and NA tubes showing no agglutination for 30 minutes at 37°C water
bath. If LISS is used, incubate for 10 minutes.
2. Centrifuge for 15 seconds at 3,400 rpm.
3. Gently dislodge the cell button and observe for hemagglutination or hemolysis.
4. If no agglutination or hemolysis is observed in MA and NA, proceed to the AHG PHASE.

C. Antihuman Globulin Phase/Coombs’ Phase (AHG)
1. Wash cells of tubes MA and NA with NSS for 3 times.
2. Decant NSS completely after the last wash.
3. Add 2 drops of AHG sera.
4. Centrifuge for 15 seconds at 3,400 rpm.
5. Dislodge the cell button and observe for hemagglutination or hemolysis.
6. Add 1 drop of Coombs’ check cells to tubes showing no agglutination.

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Interpretation

COMPATIBLE – No agglutination or hemolysis in all tubes.
INCOMPATIBLE – Presence of agglutination or hemolysis in any tubes.

GUIDE IN THE INTERPRETATION OF CROSSMATCHING TEST RESULTS:

CROSSMATCH
CASE PROCEDURAL PHASE INTERPRETATION
MAJOR MINOR
Protein Phase 0 0 Blood is compatible both in major
1 Thermo Phase 0 0 and minor crossmatch. Blood is
AHG Phase 0 0 safe for transfusion.

Protein Phase + 0 Blood is incompatible in major

crossmatch but compatible in
2 Thermo Phase 0
minor crossmatch. Blood is not safe
AHG Phase 0 for transfusion.
Protein Phase + + Blood is incompatible both in
3 Thermo Phase major and minor crossmatch. Blood
AHG Phase is not safe for transfusion.

Protein Phase 0 + Blood is compatible in major

crossmatch but incompatible in
4 Thermo Phase 0
minor crossmatch. Blood can be
AHG Phase 0 transfused but with caution.

[ + ] agglutination; [ 0 ] no agglutination

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Laboratory Activity No. 8
MLSIMHE23
BROAD SPECTRUM COMPATIBILITY TESTING

Observation

Draw and color your results. Label the tubes completely then interpret and grade the
reactions. If performed, record the result after the addition of check cells.

Unknown Sample:_________________________

PROCEDURAL PHASE TUBE REACTIONS INTERPRETATION


M MA N NA
A. Protein Phase

MA NA
B. Thermo Phase

MA NA
C. AHG Phase

Check cells reactions (if performed): ______________________________________________



_______________________________ ______________________________
Student’s Name and Signature Instructor’s Name and Signature

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Laboratory Activity No. 8
MLSIMHE23
BROAD SPECTRUM COMPATIBILITY TESTING


Post – Laboratory Questions

1. State the significance of compatibility test in transfusion medicine.






2. What is a major crossmatch? A minor crossmatch?






3. What incompatibilities are detected in the following?
a. Protein Phase




b. Thermo Phase




c. Antihuman Globulin







_______________________________ ______________________________
Student’s Name and Signature Instructor’s Name and Signature