DGGelCoombs-3.

0-IFU-en

DG Gel Coombs
Anti-Human Globulin. For in vitro diagnostic use. For professional use only.

INTENDED USE
The DG Gel Coombs card is used in gel technique for Direct and Indirect Antiglobulin Tests. These tests permit the detection of
sensitized red blood cells, screening and identification of unexpected antibodies, the determination of blood compatibility, red
blood cell typing and antibody titration in human blood samples.
The DG Gel Coombs card is intended for transfusional or investigational practices and for prenatal and perinatal
immunohematology studies.
For use in the manual method or with automated instruments.

SUMMARY AND EXPLANATION

The Polyspecific Anti-Human Globulin reagent of the DG Gel Coombs card can be used to detect red blood cells coated with
immunoglobulin (IgG) and/or complement fractions in Direct and Indirect Antiglobulin Tests.
In the Direct Antiglobulin Test (DAT), red blood cells sensitized in vivo by immunoglobulins and/or complement fractions are
detected. In the DAT, the anti-C3d activity is important in the investigation of Autoimmune Hemolytic Anemia (AIHA).
The Indirect Antiglobulin Test (IAT) is used for:
• The screening and identification of clinically significant antibodies present in the serum or plasma by in vitro sensitization
of red blood cells. An optional autocontrol test helps to distinguish autoantibodies from alloantibodies.
• The Antiglobulin Crossmatch Test (CT), to determine compatibility between donor and patient blood.
• Red blood cell typing with antisera reagents.
• ∆ Titration of antibodies against red blood cell antigens. In this case, the serum or plasma sample should be diluted in the
appropriate buffer (Grifols diluent) in order to prepare the set of dilutions before performing the Indirect Antiglobulin Test.
For investigational purposes, Coombs/Papain technique can be used to increase the sensitivity of some specific weak antibodies.
It is important to note that when using DG Gel Coombs cards for IAT, washing procedures are not required since red cell suspension
is added to the microtube before the plasma/serum, creating a barrier over the gel suspension that prevents neutralization of the
anti-human globulin by serum IgG proteins.

PRINCIPLE OF THE TEST

The principle of the test is based on the gel technique for detecting red blood cell agglutination reactions.1 DG Gel Coombs cards
are composed of eight microtubes prefilled with a buffered gel solution containing a mixture of polyclonal anti-human globulin
(anti-IgG) and monoclonal anti-C3d antibodies.
The agglutination occurs when the red blood cells sensitized in vivo or in vitro by human IgG antibodies or complement fraction
react with the anti-human globulin antibodies present in the gel solution. The gel column acts as a filter that traps agglutinated red
blood cells at the top of or along the column when they pass through the gel during centrifugation of the card. Unagglutinated
red blood cells reach the bottom of the microtube forming a pellet.
Each card is uniquely identified by a barcode. The microtubes are protected by aluminium foil.

REAGENTS
Each microtube on the DG Gel Coombs card contains a buffered gel solution containing preservative and antibodies. All microtubes
contain sodium azide (NaN3) as a preservative at a final concentration of 0.09%.
The microtubes are identified on the front label of the card:
• AHG microtubes: Polyspecific anti-human globulin in a buffered Low Ionic Strength Solution (LISS). Mixture of rabbit polyclonal
anti-IgG and murine monoclonal anti-C3d antibodies (IgM antibodies, clone 12011D10). These microtubes are green.

1/9
 DGGelCoombs-3.0-IFU-en

DG Gel Coombs

Warnings and precautions

• The results alone do not constitute a clinical diagnosis; results must be evaluated along with the patient’s clinical information.
• The product should only be used by qualified personnel.
• All products containing animal-derived material and human blood samples should be handled as if they were potentially
capable of transmitting infectious diseases.
• Once used, dispose of the product in containers for biological waste according to local, state and national regulations.
• In manual method, use disposable plastic pipette tips in order to avoid cross-contamination with samples and reagents.
• The Grifols centrifuge for DG Gel cards must be used. Use of any other centrifuge may lead to incorrect test results.

Storage and stability

• Store upright (as indicated by the two arrows on the outer packaging) with seal intact at 2-25 ºC.
• Do not freeze.
• Do not expose cards to excessive heat, air conditioning sources or ventilation outlets.
• Do not use beyond the expiration date.
• Do not use the cards if you identify incorrect temperature conditions during storage or shipment.

SPECIMEN COLLECTION AND PREPARATION

Blood samples collected in EDTA, sodium citrate or heparin should be used. No special preparation of the sample is required
prior to specimen collection. Collect samples following general blood sampling guidelines or local laboratory procedures.2,3
The recommended centrifugation range is 1000-2000 x g for 5 minutes.
Samples should be tested as soon as possible.
• For the Indirect Antiglobulin Test (IAT), use serum or plasma. Frozen samples stored up to 5 years at -20 ºC or colder may
be used after thawing. If necessary, samples stored at 2-8 ºC can be used up to 7 days after collection. If the patient has
been pregnant or transfused within the previous three months, samples stored at 2-8 ºC should be used within 72 hours
after collection.
• For the Direct Antiglobulin Test (DAT), crossmatch tests and autocontrol, use red blood cells. If necessary, samples stored
at 2-8 ºC can be used up to 7 days after collection for the crossmatch test and autocontrol, but sample storage of less than
48 hours is recommended for the DAT. If the patient has been pregnant or transfused within the previous three months,
samples stored at 2-8 ºC should be used within 72 hours after collection for crossmatch test and autocontrol.
• Red blood cells from bags collected in CPD, CPD-A or SAG-Mannitol can also be used until the expiration date indicated
on the bag’s label. If red blood cells from a bag segment are used, washing them with physiological saline solution before
preparing the suspension is recommended.
• For red blood cell typing, follow the Instructions for Use for the antisera reagent used.

PROCEDURE
Observable indications
Inspect the condition of the cards before use.
Do not use the card if any of the following is observed:
• Microbiological contamination, alterations or changes in color, or other artifacts.
• Trapped bubbles in the gel, cracked gel or gel with fissures, drying gel, or gel without a visible fine line of supernatant.
• Damaged or opened aluminium foil.
• Dispersed drops at the top of the microtube. In this case, the card should be centrifuged with the Grifols centrifuge for
gel cards before use. If after one centrifugation the drops do not descend, the card should not be used.

2/9
 DGGelCoombs-3.0-IFU-en

DG Gel Coombs

Material provided
DG Gel Coombs cards are ready to use.

Code Product Designation Packaging Profile

210342 DG Gel Coombs 2 x 25 Cards 8x (AHG)

Material required but not provided

Manual Method
• 10 μL, 25 μL, 50 μL and 1 mL automatic pipettes.
• Disposable pipette tips.
• Glass or plastic test tubes.
• Grifols diluent.
• Grifols incubator for DG Gel cards.
• Grifols centrifuge for DG Gel cards.
• Grifols Reagent Red Blood Cells 0.8%.
• Antisera reagents.
• Grifols reader for DG Gel cards (optional).
Fully Automated Methods
• Grifols diluent.
• Grifols Reagent Red Blood Cells 0.8%.
• Antisera reagents.
• DG Fluid A and DG Fluid B.
• Grifols automated instruments.

Test procedure
1. Allow DG Gel Coombs cards, additional reagents and the samples to reach room temperature (18-25 ºC).
Note: For fully automated instruments, skip the following steps and refer to the instrument’s Instructions for Use.
2. Identify the cards to be used and the samples to be tested.

• Direct Antiglobulin Test (DAT)

3. Prepare a 1% red blood cell suspension in Grifols diluent (10 μL of packed red blood cells in 1 mL of Grifols diluent).
4. Carefully remove the aluminium foil from the entire gel card or the microtubes to be used for testing.
Note: Use the microtubes immediately once the aluminium foil has been removed.
5. Ensure the homogeneity of the 1% red blood cell suspension before use.
6. Dispense 50 μL of the 1% red blood cell suspension into the corresponding microtube.
7. Centrifuge the gel card in the Grifols centrifuge.
8. After centrifugation, read the results visually or use a Grifols reader.

• Screening and/or Identification of ∆ antibodies test (IAT)

3. Thoroughly mix the vials of Reagent Red Blood Cells to ensure a homogeneous suspension of the red blood cells before use.
Note: Use Papainized Reagent Red Blood Cells in the Coombs/Papain technique.
4. Carefully remove the aluminium foil from the entire gel card or the microtubes to be used for testing.
Note: Use the microtubes immediately once the aluminium foil has been removed.
5. Dispense 50 μL of Reagent Red Blood Cells into the microtubes.
Note: Use Grifols Reagent Red Blood Cells for reverse grouping to detect ABO antibodies.
6. Add 25 μL (for non-ABO antibodies) or 50 μL (for anti-A and anti-B antibodies) of serum or plasma to the same microtubes.
7. Incubate 15 minutes at 37 ºC using the Grifols incubator.
8. Centrifuge the gel card in the Grifols centrifuge.
9. After centrifugation, read the results visually or use a Grifols reader.

3/9
 DGGelCoombs-3.0-IFU-en

DG Gel Coombs

• Crossmatch test (IAT) and Autocontrol

3. Prepare a 1% red blood cell suspension in Grifols diluent (10 μL of packed red blood cells in 1 mL of Grifols diluent). Ensure
the homogeneity of the 1% red blood cells suspension.
Note: For autocontrol, use the patient’s own red blood cells and plasma or serum.
4. Carefully remove the aluminium foil from the entire gel card or the microtubes to be used for testing.
Note: Use the microtubes immediately once the aluminium foil has been removed.
5. Dispense 50 μL of the 1% red blood cells suspension into the microtube.
6. Add 25 μL of serum or plasma to the same microtube.
7. Incubate for 15 minutes at 37 ºC using the Grifols incubator.
8. Centrifuge the gel card in the Grifols centrifuge.
9. After centrifugation, read the results visually or use a Grifols reader.

• Antibody titration test (IAT)

3. Use Grifols diluent to perform serial dilution of the antibody in a test tube (100 μL of plasma or serum in 100 μL of
Grifols diluent).
Note: Dilution set should be used immediately. If necessary, it may be used up to 1 hour after preparation.
4. Select the Reagent Red Blood Cells with the same specificity as the antibody presented in the sample. Thoroughly mix
the vial of Reagent Red Blood Cells to ensure a homogeneous suspension of the red blood cells before use.
Note: If it is not possible to use Reagent Red Blood Cells, prepare a 1% red blood cell suspension in Grifols diluent.
5. Carefully remove the aluminium foil from the entire gel card or the microtubes to be used for testing.
Note: Use the microtubes immediately once the aluminium foil has been removed.
6. Dispense 50 μL of Reagent Red Blood Cells into the microtubes.
7. Add 25 μL (for unexpected antibodies) or 50 μL (for anti-A and anti-B antibodies) of each serum/plasma dilution to the
same microtubes.
Note: The antibody titration test could be executed pipetting only one dilution (single dilution titration test).
8. Incubate for 15 minutes at 37 ºC using the Grifols incubator.
9. Centrifuge the gel card in the Grifols centrifuge.
10. After centrifugation, read the results visually or use a Grifols reader.

• Red blood cell typing test (IAT)

Follow the Instructions for Use for the antisera reagent used.

Stability of the results

Do not leave processed cards in a horizontal position. If necessary, a delayed reading can be performed up to 24 hours after
processing the cards if they are kept in an upright position, refrigerated (2-8 ºC), and sealed with laboratory film to prevent
evaporation of the supernatant.
Do not read the results of microtubes that have been centrifuged more than once.
Note: In the 24-hour delayed reading of processed cards with weak positive samples, a loss in agglutination intensity may
be observed.

Quality control
The inclusion of positive and negative controls according to the laboratory’s policies is recommended. If an unexpected control
result is obtained, a complete assessment of the instrument, reagents and materials used should be made.

4/9
 DGGelCoombs-3.0-IFU-en

DG Gel Coombs

RESULTS
Report the results as an agglutination grade, absence of agglutination, hemolysis or Double Population:

Negative Well-defined pellet of unagglutinated red blood cells at the bottom of the gel column and no visible
-
agglutinated cells or hemolysis in the rest of the gel column.
Positive Barely visible small-sized clumps of agglutinated cells in the lower part of the gel column and a pellet of
+/-
unagglutinated cells at the bottom.
Some small-sized clumps of agglutinated cells, most frequently in the lower half of the gel column. A
1+
small pellet may also be observed at the bottom of the gel column.
Small or medium-sized clumps of agglutinated cells throughout the gel column. A few unagglutinated
2+
cells may be visible at the bottom of the gel column.
3+ Medium-sized clumps of agglutinated cells in the upper half of the gel column.
A well-defined band of agglutinated red blood cells in the top part of the gel column. A few agglutinated
4+
cells may be visible below the band.
Double A band of red blood cells in the top part of the gel or dispersed throughout the gel column and a pellet of
DP
Population unagglutinated cells at the bottom of the microtube.
Hemolysis Hemolysis in the microtube with very few or no red blood cells in the gel column. Annotate if hemolysis is
H
present in the microtube but not in the sample.

– +/- 1+ 2+ 3+ 4+ DP H
Figure 1. Example of agglutination grades

Interpretation of the results

• Direct Antiglobulin Test (DAT)
- Negative reaction: Indicates the absence of detectable IgG antibodies or C3d complement component on
the red blood cells.
- Positive reaction: Indicates that the patient’s red blood cells are sensitized (red blood cells coated with
IgG antibodies and/or C3d).

• Antibody screening, antibody identification tests (IAT)

- Negative screening: Indicates the absence of detectable ∆ antibodies in the patient’s or donor’s serum or plasma.
- Positive screening: Indicates the presence of ∆ antibodies in the patient’s or donor’s serum or plasma against one
or more antigens present on the Reagent Red Blood Cells.
In case of positive screening, an identification panel should be performed to identify the
antibody present in the plasma or serum sample.
Note: The autocontrol should be negative. If the autocontrol is positive, it may indicate the presence of autoantibody in the
sample or a non-specific reaction.
The antigen table provided with the Reagent Red Blood Cells should be used to identify the antibody present in the sample.

5/9
 DGGelCoombs-3.0-IFU-en

DG Gel Coombs

• Crossmatch test (IAT)

- Negative reaction: Indicates compatibility of the donor blood with the recipient.
- Positive reaction: Indicates incompatibility of the donor blood with the recipient due to the presence of
antibodies directed against antigens on the donor’s red blood cells. Further investigation
to identify the antibody specificity should be performed.
Note: The autocontrol should be negative. If the autocontrol is positive, it may indicate the presence of autoantibody
in the sample or a non-specific reaction.

• Antibody titration test (IAT)

The titer is the reciprocal of the highest dilution that produces a positive reaction (≥1+) followed by a negative reaction
(e.g., in a 1:16 dilution, the titer = 16).4

• Red blood cell typing test (IAT)

Follow the Instructions for Use for the antisera reagents used.
Notes:
1. The checking or investigation of any discrepancy in the results is recommended.
2. The observation of complete or partial hemolysis (pinkish supernatant and/or gel column) in microtubes should be interpreted
as a positive result once it has been verified that this is not due to a problem with the collection and/or handling of the sample.
Hemolysis of red blood cells usually indicates the presence of high titers of alloantibodies or autoantibodies. The presence of
A and B antibodies may also cause hemolytic disease of the newborn due to ABO incompatibility.
3. Occasionally, red blood cells may be retained in the incubation chamber with positive 4+ samples; this does not interfere with
the reading of results.

LIMITATIONS OF THE PROCEDURE

1. Grossly hemolyzed, cloudy, or contaminated samples or samples containing a clot may give false positive or false negative
results.
2. Aged or hemolyzed specimens may cause weaker reactions compared with those obtained with a fresh sample.
3. Samples with high-potency antibodies may coat the red blood cells completely, causing spontaneous agglutination.4
4. Abnormal concentrations of serum proteins, the presence of macromolecular solutions in the serum/plasma, or the presence
of Wharton’s jelly in cord blood samples may cause non-specific agglutination of the red blood cells. Washing the red blood
cells before performing the test is recommended.4
5. The presence of some drugs or dextran solutions, or the remains of silicone gel from the extraction tube in the sample may
cause a positive result in the Direct Antiglobulin Test.4
6. The presence of high concentrations of immunoglobulins and other serum proteins in the sample may lead to a false positive
result in antiglobulin tests.4
7. False positive results may occur if antibodies to components of the preservative solution are present in the sample tested.
8. Antibody activity may decrease in the elderly, infants or persons with disease.
9. The use of Papainized Reagent Red Blood Cells may lead to non-specific agglutinations in Indirect Antiglobulin Tests.5,6
10. If plasma is used, complement-dependent hemolytic reactions may not be detected.
11. If poorly anti-coagulated plasma or incompletely clotted serum is used, fibrin residues may trap unagglutinated red blood cells
at the top of the gel, appearing as a pinkish or reddish layer. Although the results may be correctly interpreted, in a negative
reaction, the false appearance of a Double Population may lead to misinterpretation. In the case of incompletely clotted serum
samples, re-clotting the serum for 10 minutes at 37 ºC, centrifuging, and repeating the test are recommended.4
12. On occasion, unagglutinated red blood cells may be retained somewhere in the gel column, appearing as a very minute red
dot or fleck; this nonspecific retention, however, should not interfere with the interpretation of the result.
13. In the Direct Antiglobulin Test, not all negative reactions indicate the absence of hemolytic disease of the newborn, especially
in cases where ABO incompatibility is suspected.
14. In the Direct Antiglobulin Test, not all positive reactions indicate that clinically significant antibodies are present. Specific
anti-IgG reagent and elution techniques may be used for additional investigation of positive results.

6/9
 DGGelCoombs-3.0-IFU-en

DG Gel Coombs

15. A false positive result in the Direct Antiglobulin Test may be due to the complement attached to red blood cells in specimens
collected from infusion lines used to administer dextrose-containing solutions4 or in specimens collected in tubes containing
silicone gel.4
16. Nonspecifically adsorbed proteins (e.g., high-dose intravenous immune globulin, multiple myeloma, autoimmune disorders
and other diseases associated with elevated serum globulin) and the modification of the red blood cell membrane by some
drugs may cause a positive result in the Direct Antiglobulin Test.4
17. If an unexpected result is obtained in the Direct Antiglobulin Test, washing the red blood cells with physiological saline
solution, preparing a new suspension of the washed red blood cells, and repeating the test are recommended.
18. Red blood cell samples with a positive Direct Antiglobulin Test result should not be used for Indirect Antiglobulin testing.
19. The polyclonal anti-IgG used in the DG Gel Coombs card is not heavy chain specific, and thus may also react weakly with
red blood cells coated with IgA and/or IgM molecules due to anti-light chain activity.
20. For anti-A and anti-B titration studies, 25 μL of plasma/serum dilution may be used in the test procedure. Each laboratory
should establish the antibody titer threshold using the titration procedure, taking into account clinical findings and laboratory
data to ensure meaningful interpretation based on its own titration values.
21. No single method is able to detect all unexpected antibodies. The optimum reaction conditions (e.g., sample volume,
incubation time) may vary for different antibody specificities. For screening and identification of unexpected antibodies,
crossmatch, autocontrol, and titration tests, increasing the volume of serum or plasma from 25 μL to 50 μL is acceptable.
This variation in the concentration of antibodies brings the antigen/antibody ratio down and may improve the detection of
antibodies at very low concentrations.4
22. If antibody detection test is used for antibody single dilution titration studies, the laboratory should establish the antibody titer
threshold using the procedure with clinical findings and laboratory data to ensure meaningful interpretation based on its own
titration values.
23. If there are antibodies against high-incidence antigens or multiple antibodies in the sample, all Reagent Red Blood Cells may
be agglutinated.
24. If a patient has been recently transfused, results obtained with the autologous control must be interpreted carefully, since
alloantibody capable of reacting with circulating donor red blood cells may lead to agglutination of the autologous control.
25. In red blood cell typing, DP (Double Population) events should be interpreted cautiously. Not all DP situations are detected.
Additional information on patient history and additional testing will be necessary to reach a determination. Transfused patients
or those who have undergone bone marrow transplantation may present this pattern.4
26. In the detection of weak agglutination reactions of the ABO system, performing the Indirect Antiglobulin Test at 37 ºC using
gel or glass sphere techniques has been reported to show a lower level of sensitivity than results obtained with the tube
technique.7
27. In antibody detection using enzymatic techniques, some enzyme-treated red blood cells may be retained at the end of the
column when the level of supernatant is low (around 1 mm), leading to a false weak positive result. Enzyme techniques are
difficult to standardize and appropriate controls should be used with enzyme-treated red blood cells.
28. The use of different volumes and/or other concentrations of red blood cell suspensions than those indicated in the Instructions
for Use, or the use of a non-Grifols diluent may modify the reaction and produce incorrect test results (e.g., false positives or
false negatives).

7/9
 DGGelCoombs-3.0-IFU-en

DG Gel Coombs

SPECIFIC PERFORMANCE CHARACTERISTICS

Method comparison
For the manual method, the performance of the reagent was confirmed against other CE-marked products in a method comparison
study. The percentages of agreement and their lower confidence limits estimated with a 95% confidence interval are indicated on
the table below:

Negative Percent Agreement Positive Percent Agreement

Antibody Technique
(Lower 95% CI) (Lower 95% CI)
98.6% 98.9%
DAT(a)
(97.2%) (94.9%)
AHG 99.9% 100.0%
IAT(b) - Investigation
(Anti-IgG, -C3d) (99.9%) (98.0%)
100.0% 100.0%
IAT(b) - Crossmatch
(95.1%) (95.1%)
(a) DAT: Direct Antiglobulin Test
(b) IAT: Indirect Antiglobulin Test

Percentages of agreement only indicate agreement between the Diagnostic Grifols reagents and other CE-marked products
and do not indicate which reagent obtained the correct result(s).
The performance of the antibody titration test using DG Gel cards was evaluated obtaining the expected titer, showing that
DG Gel cards can be used for titration of unexpected and natural antibodies of human blood samples.
For further information about the performance data for red blood cell typing, please refer to the Instructions for Use of the
antisera reagent.

Precision
The precision of the reagents present in the DG Gel Coombs card was evaluated, including repeatability, inter-lot reproducibility
and intra-laboratory reproducibility tests. No discrepant results were obtained, and differences between agglutination intensities
in positive samples did not exceed 1+ in all assays.

BIBLIOGRAPHY
1. Lapierre Y, et al. The gel test: a new way to detect red cells antigen-antibody reactions. Transfusion, 30: 109-113, 1990.
2. CLSI GP41: Collection of diagnostic venous blood specimens; Approved Standard, 7th edition, 2017.
3. CLSI GP44-A4: Procedures for the handling and processing of blood specimens for common laboratory tests; Approved
Guideline, 4th edition, 2010.
4. Technical Manual, 20th edition, American Association of Blood Banks, Bethesda, Maryland, 2020.
5. John Libbey, 1st edition, Les analyses immunohématologiques et leurs applications cliniques / Clinical applications of
immunohematology assays. Paris; 69, 2011.
6. Mollison’s Blood Transfusion in Clinical Medicine. 11th edition. Bristol, UK; 317, 2005.
7. Phillips P, et al. An explanation and the clinical significance of the failure of microcolumn tests to detect weak ABO and
other antibodies. Transfusion Medicine, 7: 47-53, 1997.

8/9
 DGGelCoombs-3.0-IFU-en

DG Gel Coombs

Manufactured by:
Diagnostic Grifols, S.A.
Passeig Fluvial 24, 08150 Parets del Vallès (Barcelona), Spain
Tel. 935 71 04 00
www.grifols.com
Date of issue: July 2023
Serious incidents occurring in relation to the device in the European Union should be reported to the manufacturer and the competent authority
of the Member State in which the user and/or the patient is established.

The Summary of Safety and Performance (SSP) will be available on https://ec.europa.eu/tools/eudamed once EUDAMED is operational.

This document is available in several languages. In the event of doubts or discrepancies, the wording in the master document in English shall
take precedence.

SYMBOLS KEY
One or more of these symbols may be used in the labeling/packaging of this product.

In vitro diagnostic Box raw material

medical device Consult instructions for use This way up (paperboard)

Batch code Catalogue number Fragile, handle with care

Use by Cards Keep dry

Recycled and recyclable

Temperature limitation Manufacturer
100%

packaging

REVISION HISTORY

Revision Date Revisions*

2021-07 Original issue
2021-12 • Limitation “The use of Papainized Reagent Red Blood Cells may lead to non-specific agglutinations
in Indirect Antiglobulin Tests” has been added in Section - Limitations of the procedure.
• The following references have been added in Section - Bibliography: John Libbey, 1st edition,
Les analyses immunohématologiques et leurs applications cliniques / Clinical applications of
immunohematology assays. Paris; 69, 2011 and Mollison’s Blood Transfusion in Clinical Medicine.
11th edition. Bristol, UK; 317, 2005.
• Create a new Revision history Section.
2023-07 • Additional information has been included in Section - Summary and explanation to improve the
comprehension of the Indirect Antiglobulin Test (IAT) uses.
• A new note has been added in step 3 and 5 of the “Screening and/or Identification of antibodies
test (IAT)” procedure described in Section - Test procedure.
• The serum/plasma volume to be used for the investigation of ABO antibodies has been included
in Section - Test procedure.
• A new note has been added in step 7 of the “Antibody titration test (IAT)” procedure described
in Section - Test procedure.
• Limitations nº 22 and nº 27 have been added in Section - Limitations of the procedure.
• Minor editorial corrections have been applied in Section - Summary and explanation, Section -
Test procedure and Section - Interpretation of the results.

* Note: Changes with respect to the previous revision are shaded in grey. Deletion of text is indicated as ∆.

9/9