Ra Physicaltherapy Manila Feb2019

COMPATIBILITY TESTING – GEL CARD TEST ANTI-IgG
PRINCIPLE
The crossmatch test is used to detect the presence of blood group antibodies in a recipient's serum/plasma to antigens on
donor red blood cells. In this test, the donor red blood cells in a hypotonic buffered saline solution are combined with
patient serum/plasma to allow antigen/antibody interaction in the upper chamber of the microtube. This results in
promoting antibody uptake. The detection of this antibody occurs when the sensitized red blood cells react with the Anti-
IgG gel in the microtube during centrifugation.
SPECIMEN
Draw one Edta tube. Spin for 5 minutes at 3,400 rpm.
No special preparation of the patient is required prior to specimen collection. Blood should be collected by approved
techniques. The anticoagulated sample must be drawn within 24 hours of testing.
REAGENTS
MTS Diluent 2 , a hypotonic buffered saline solution
MTS Anti-IgG Card , Anti-IgG (Rabbit) suspended in gel
Do not use beyond expiration date. Store at 2 to 25o C. Use at room temperature (18 to 25 C).
QUALITY CONTROL
To recognize reagent deterioration, the reagents must be tested daily with appropriate controls. See Quality Control
procedure in this manual. MTS Diluent 2 must be visually checked to ensure that the liquid is not discolored, turbid or
showing any signs of bacterial contamination. Also a known positive and negative cell should be diluted with each lot of the
diluent and tested by the IAT and/or DAT procedure with the MTS Gel Card System being used.
PROCEDURE
Donor Unit Cell Preparation

1. Label test tubes with the appropriate donor unit numbers.
2. In separate labeled tubes place 1 mL of MTS Diluent 2 . Add 10 µL of the packed red blood cells from each donor unit.
3. Mix gently. Final cell suspension should be approximately 0.8%.
Compatibility Test Procedure

1. Upon orders for blood, review the blood bank recipient card file and/or computer blood bank history for previous
testing. Interpretation of previous results should be compared with current interpretations and any discrepancies
must be resolved.
2. Prior to or in conjunction with the crossmatch, an ABO and Rh type and antibody screen must be performed on the
patient (recipient) sample to assure that type specific blood is given. If an antibody is detected, the blood selected
for crossmatch should be negative for the corresponding antigen. (See appropriate procedures in this manual). Be
sure that the patient name and identification number agree on the tubes of blood and requisition slip. If there is any
discrepancy, redraw the patient. Autologous units will be processed the same as any other unit. The unit will be
destroyed if not used.
3. Label the MTS Anti-IgG Card with the appropriate patient identification and test information.
4. Remove the foil seal from the card.
5. Using an appropriate pipette, add 50 µL of each donor unit red blood cell suspension to the correct microtube.
6. Using an appropriate pipette, add 25 µL of serum or plasma to the correct micro tube(s).
7. Incubate at 37 C for 15 - 30 minutes.
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8. Centrifuge the gel card for 10 minutes.
9. Read the front and the back of each microtube macroscopically and record reactions as described in the
INTERPRETATION section below.
INTERPRETATION
Negative – a complete sedimentation of all red cells on the bottom of the microtube.
Strong positive – cells remain suspended on the top of the gel.
Weak positive – cells are dispersed throughout the gel.
Mixed cell reactions may be recognized as a layer of positive cells at the top of the gel accompanied by a button of negative
cells at the bottom of the microtube.
Clots, particulates or other artifacts may cause some cells to be entrapped at the top of the gel which may cause an
anomalous result in a negative test.
A few cells may form a button at the bottom of the microtube in some positive reactions.
Refer to ID-Micro Typing System Interpretation Guide for additional grading information.
RESULTS
Hemolysis or agglutination of a crossmatch in the gel card indicates the presence of an antibody directed against the
corresponding antigen which is present on the donor cells. This result is interpreted as INCOMPATIBLE.
No agglutination or hemolysis of a crossmatch is a negative test result and indicates the absence of an antigen/antibody
reaction. This result is interpreted as COMPATIBLE.
If any part of the crossmatch is incompatible, the problem must be resolved before the blood is issued. (See the
“Antibody Identification” procedure in this manual.) Only group-compatible, Rh-specific blood is to be used for
crossmatching. Only crossmatch-compatible blood is to be used for transfusion. When a recipient has been identified
with an antibody, only antigen-negative, crossmatch-compatible blood is used for transfusion. Patients will receive
irradiated, CMV negative and Hemoglobin S negative units when ordered by the physician.
COMMENTS
Interpretation of mixed-field reactions must be done with caution. The presence of fibrin, clots or particulates may result in
some cells layering at the top of the gel. Patient clinical information should be reviewed before concluding a test is mixed-
field.
LIMITATIONS
Antibodies with levels below the threshold level of detection may not be detected by this test.
False-positive results may occur if antibodies to components of the preservative solution are present in the serum tested.
Significant variations in red blood cell suspensions may result in false-positive or false-negative reactions.
Anomalous results may be caused by fresh serum, fibrin or particulate matter in serum or plasma, or red cells that stick to
the sides of the microtube. Anomalous results with fresh serum (i.e. a line of red cells on top of the gel) may be minimized
by the use of EDTA plasma.
Adherence to the test procedure is critical to test performance.
REFERENCES
AABB Technical Manual, 14th Edition, 2002; pp. 384-391.
Technical manual. 11th ed. Bethesda, MD: American Association of Blood Banks, 1993:314-6, 313-30.
Package insert: Anti-Human Globulin Anti-IgG (Rabbit) MTS Anti-IgG Card . Pompano Beach, FL: Micro Typing Systems, Inc.
Package insert: MTS Diluent 2 Red Blood Cell Diluent. Pompano Beach, FL: Micro Typing Systems, Inc.
Malyska H, Weiland D. The gel test. Laboratory Medicine 1994;25:81-5.
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Consortium for Healthcare Education Online project material by CHEO

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COMPATIBILITY TESTING – GEL CARD TEST ANTI-IgG

Donor Unit Cell Preparation

Compatibility Test Procedure

Consortium for Healthcare Education Online project material by CHEO

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