Document No.

:
TGMCI-03-LAB-148-00

Address: Purok Cafe, Visayan Village, Tagum City

BLOOD BANK
Page: 1 of 7
(CROSSMATCHING – GEL METHOD)

REVISION HISTORY

REASON EFFECTIVITY
REG. REVISION PAGE PREPARED REVIEWED APPROVED
DCN FOR DATE
DATE STATUS AFFECTED BY BY BY
REVISION

Creation Kristine Joy John Patrick Glenn A.

07-18-23 00 ALL 07-18-23
C.Ochavillo, C. Padilla, Pono, MD
RMT MD, FPSP Medical
Head Director
Pathologist

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reproduction is strictly prohibited unless otherwise permitted by ACE-TGMCI Management.
 Document No.:
TGMCI-03-LAB-148-00

Address: Purok Cafe, Visayan Village, Tagum City

BLOOD BANK
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(CROSSMATCHING – GEL METHOD)

1.0 PURPOSE OF EXAMINATION

1.1. The test demonstrates incompatibility and clinically significant antibodies
to red cell antigens.

2.0 PRINCIPLE OF THE PROCEDURE

2.1. Hemeagglutination Antigen test in a Gel Medium.The purpose of the
test is to detect in vitro sensitization between serum and red cells.

3.0 PERFORMANCE SPECIFICATION

3.1. Saline techniques are used to detect antibodies that react predominantly
at 4 °C or at 18 – 25 °C (room temperature), such as anti-M, -N, -P1, -Lea,
-Leb, -I. They are also used to define hemolytic anemia associated with
cold antibodies.
3.2. Enzyme techniques are useful when increased sensitivity in antibody
screening is desired. They enhance the reactions of certain antibodies,
notably in the Rh, Kell and Kidd systems. Since proteolytic enzymes
usually destroy some antigens such as M (MNS1), N (MNS2), S (MNS3),
Fy(a) (FY1), Fy(b) (FY2), an enzyme technique should never be the only
method employed.
3.3. It is not considered necessary to include the room temperature saline test
in routine antibody screening procedures, but the saline test at 4 °C can be
used to detect cold agglutinins
3.4. ID-Card ˝LISS/Coombs˝ with 6 microtubes containing polyspecific AHG
(rabbit anti-IgG and monoclonal anti-C3d, cell line C139-9) within the gel
matrix.

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reproduction is strictly prohibited unless otherwise permitted by ACE-TGMCI Management.
 Document No.:
TGMCI-03-LAB-148-00

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BLOOD BANK
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(CROSSMATCHING – GEL METHOD)

1.0 PRIMARY SAMPLE SYSTEM

1.1. 0.8% Red Cell Suspension of Patient
1.2. 0.8% Red Cell Suspension of Donor
2.0 TYPE OF CONTAINER AND ADDITIVES
2.1. Whole blood in EDTA Tube
2.2. Tubing obtained originally attached to the blood container.

3.0 REQUIRED EQUIPMENTS AND REAGENTS

3.1. ID Diluent 2: LISS
3.2. Pipettor
3.3. Dispenser
3.4. Tips
3.5. Working Table
3.6. Suspension tubes
3.7. Centrifuge
3.8. Incubator
3.9. Timer

4.0 CALIBRATION PROCEDURES (N/A)

5.0 PROCEDURAL STEPS

5.1. Preparation of 0.8% Red Cell suspension

5.1.1. Allow the diluent to reach room temperature before use.
5.1.1.1. Dispense 1.0 ml of ID-Diluent 2 into a clean tube.
5.1.1.2. Add 10 μl of packed red cells, mix gently. Where
whole blood directly from a segment of the blood
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 Document No.:
TGMCI-03-LAB-148-00

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BLOOD BANK
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(CROSSMATCHING – GEL METHOD)

bag is used, add 20 μl of blood to the 1.0 ml of ID-

Diluent 2.
NOTE: The cell suspension may be used immediately.

5.2. With NaCl (Saline Phase)

5.2.1. Identify the ID-Card with the unique patient or donor
number/details as appropriate.
5.2.2. Remove the aluminium foil from as many microtubes as required
by holding the ID card in the upright position.
5.2.3. Pipette 50 μl of the donor’s red cell suspension to the appropriate
microtubes.
5.2.4. When an autocontrol is to be included, add 50 μl of the patient’s
red cell suspension to the appropriate microtube.
5.2.5. Add 25 μl of the patient’s plasma or serum to each microtube.
5.2.6. Incubate the ID-Card for 15 minutes at room temperature (18–25
°C).
5.2.7. Centrifuge the ID-Card for 10 minutes in the ID-Centrifuge.
5.2.8. Read and record the results.

5.3. With LISS Coombs (AHG & Thermal Phase)

5.3.1. Identify the appropriate microtubes of the ID-Card ˝LISS/Coombs˝
with recipient’s and donor’s name or number.
5.3.2. Remove the aluminium foil from as many microtubes as required
by holding the ID-Card in the upright position.
5.3.3. Pipette 50 μl of the donor red cell suspensions to the appropriate
microtubes.
5.3.4. For the autocontrol, pipette 50 μl of the patient’s own red cell
suspension to the appropriate microtube.
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 Document No.:
TGMCI-03-LAB-148-00

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BLOOD BANK
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(CROSSMATCHING – GEL METHOD)

5.3.5. Add 25 μl of the patient’s plasma or serum to each microtube.

5.3.6. Incubate the ID-Card for 15 minutes at 37 °C in the ID-Incubator.
5.3.7. Centrifuge the ID-Card for 10 minutes

6.0 QUALITY CONTROL OF THE PROCEDURE (N/A)

7.0 INTERFERENCES

7.1. ID-Cards which show air bubbles or gel drops in the upper part of the
microtubes and/or the seal, must be centrifuged before use.
7.2. Certain drugs are known to cause positive reactions in anti-human globulin
procedures.
7.3. Some pathological conditions are also reported as causing positive
reactions in anti-human globulin procedures.
7.4. Cells that have become polyagglutinable, due to cryptantigen exposure e.g.
T antigen, either in vivo or in vitro may react with all human sera. Further
investigation of such reactions is required.
7.5. Bacterial or other contamination of materials used can cause false positive
or false negative results.
7.6. Fibrin residues in the red cell suspension may trap non-agglutinated cells
presenting a fine pink line on top of the gel while most of the cells are on
the bottom of the microtube after centrifugation.
7.7. Strict adherence to the procedures and recommended equipment is
essential. The equipment should be checked regularly according to GLP
procedures.

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BLOOD BANK
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(CROSSMATCHING – GEL METHOD)

7.8. Use of suspension solutions other than ID-Diluent 2 may modify the
reactions.
7.9. The use of other test cell reagents than ˝ID-DiaCell˝ or ˝ID-DiaPanel˝ may
modify the reaction patterns.
7.10. Too heavy or too weak red cell suspensions can cause aberrant results.

8.0 PRINCIPLE OF PROCEDURE FOR CALCULATING RESULTS (N/A)

9.0 BIOLOGICAL REFERENCE INTERVAL

10.0 REPORTABLE INTERVAL OF EXAM (N/A)

11.0 ALERT/CRITICAL VALUES (N/A)

12.0 INTERPRETATION

12.1. Agglutination of cells or formation of a red line on the surface of the gel
constitutes a positive test result.
12.2. Compact button of cells on the bottom of the microtubule.
12.3. A negative reaction indicates compatibility of the donor blood with the
recipient.
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reproduction is strictly prohibited unless otherwise permitted by ACE-TGMCI Management.
 Document No.:
TGMCI-03-LAB-148-00

Address: Purok Cafe, Visayan Village, Tagum City

BLOOD BANK
Page: 7 of 7
(CROSSMATCHING – GEL METHOD)

NOTE: A positive reaction indicates incompatibility of the donor blood with

the recipient, due to presence of antibodies directed against antigens on
the donor red cells. Further investigation to identify the antibody specificity
should be performed.

13.0 SAFETY PRECAUTION

13.1. Always wear personal protective equipment’s when handling samples.
13.2. All samples are treated as infectious material and should be disposed
according to protocol.

14.0 POTENTIAL CAUSES OF VARIABILITY (N/A)

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reproduction is strictly prohibited unless otherwise permitted by ACE-TGMCI Management.