NAME OF THE LAB ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,

Name Position Signature Date

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Reviewed by

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1.0 Purpose

The purpose of this procedure is to give instructions on how to perform compatibility testing in a
standard way.

2.0 Scope

This procedure is used for compatibility tests done at ,,,,,,,,,,,,,,,,, laboratory. This test is normally
done for patients who need blood transfusion.

3.0 Principle

Washed donor red cells are added to the recipient’s serum to detect the presence of antibodies
after immediate spin and incubation at 37 degrees centigrade.

4.0 Responsibility

It is the responsibility of the Head of Blood Bank to ensure effective implementation of this
procedure.

5.0 Equipment and Reagents

Equipment Materials/ Reagents

Weighing scale -Antisera anti-A, anti-B, anti-AB anti-D
Centrifuge -Sodium chloride powder
Glass test tubes, small(75×10 mm) -Distilled water
Pasteur Pipettes -Grease pencil
Microscope -Suspensions of known A,B and O cells
Water bath -IgG sensitized O cells
Glass slides -AHG
Cover slips

5.1 Preparation of reagents

Physiological saline:

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 Dissolve 9 g of sodium chloride in 1000 ml distilled water in a reagent bottle and

label with the name of reagent and date of preparation. Transfer into a dropper
bottle and label.

Sensitized O cells:

 Place 4 drops of known O positive blood in a test tube.

 Add two drops of incomplete anti-D.
 Incubate at 37 degrees centigrade in a water bath for 1 hour.
 Wash the cells 3 times by adding 5-7 ml physiological saline, centrifuging at 3000
rpm for 2-3 minutes and discarding the supernatant.
 Make a 3-5 % suspension of IgG sensitized O cells by mixing one drop of
sedimented cells in 20-25 drops of saline, holding the Pasteur pipette vertically.

6.0 Samples required

-Fresh serum and red cells from recipient.e. blood in EDTA and plain tube

- 3-5% suspension of donor red cells in physiological saline.

-Prepare a 3-5 % red cell suspension as follows:

 Transfer about 0.5 ml of patients red cells into a test tube with about 5ml physiological
saline.
 Wash the cells by centrifuging at 3000 rpm for 2-3 minutes and discarding the
supernatant in one quick motion.
 Repeat the above two steps 2 times.
 Make a 3-5% red blood cell suspension by mixing I drop of segmented cells in 20-25
drops of saline, holding the Pasteur pipette vertically.

7.0 Safety Precaution

Treat all samples as potentially infectious.

8.0 Quality Control

 Quality control of antisera and LISS/Bovine albumin is done on daily basis or whenever
new batch is opened.
 AHG is controlled every time a compatibility test is done by adding O sensitized cells.

9.0 Activity Description

1. Carry out ABO and Rh grouping on the recipient’s red blood cells.

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2. Select blood of the appropriate group for the recipient. Record the number of the donor
blood unit on your worksheet.
3. Label two tubes with the donor blood unit number, one tube as auto control, another as
S1 and another one as S2 with a grease pencil.
4. Cut the last segment of the blood donor set and transfer 2-4 drops into one tube.
5. Wash the cells three times and make a 3-5 % cell suspension as described above.
6. Into the second test tubelabeled with donor number, dispense 1 drop of washed 3-5% cell
suspension of donor red cells. Into the tube labeled auto control, dispense 1 drop of
washed 3-5% cell suspension of recipient cells and into the tubes labeled S1 and S2,
dispense screening cells S1 and S2 respectively.
7. Add 2 or 3 drops of recipient’s serum to all the 4 tubes to achieve approximately a 2:1
ratio of recipient’s serum to red cells. NB: Droppers used to dispense red cells and serum
should be of equivalent size.
NB: Treat all the 4 tubes according to steps 8 to 15
8. Centrifuge at low speed; 1000 rpm for one minute.
9. Observe supernatant for hemolysis .Then re-suspend the cell button by gently rolling the
tube between the thumb and the first finger to check for agglutination. Record results for
your immediate spin.
10. In case of no hemolysis or agglutination, add two drops of bovine albumin (or other
enhancement medium such as LISS to the tube. Mix and incubate for 30 minutes at 37
degreescentigrade. If LISS is used instead of albumin lower incubation time to 15
minutes.
11. Centrifuge as above, observe supernatant for hemolysis and re-suspend cells to check for
agglutination. Record results.
12. In case of no hemolysis or agglutination, wash the cells 3-4 times using normal saline
13. Add 2 drops of AHG and centrifuge at 1000 rpm for 1 minute.
14. Examine for hemolysis or agglutination macroscopically. Record results.
15. Add one drop of IgG-sensitized O cells to each negative test. Centrifuge and examine for
agglutination both macroscopically and microscopically by placing a drop of sediment
cells on a slide, putting a cover slip and examining using the x10 objective. Test must be
positive; otherwise results of the whole procedure are invalid and must be repeated.

10. 0 Reporting and interpretation of results

 Refer to Procedure for ABO Grouping

 No agglutination or hemolysisat the end of a compatibility testing (before controlling
with O sensitized cells) means the unit is compatible.

11.0 Limitations

 Patients who have had multiple transfusions in the recent past may end up with
distorted blood groups. Always check the history of the patient.

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 No current testing procedure can guarantee the fate of a unit of blood that is to be
transfused. Even a compatible cross match cannot guarantee that the transfused red
cells will survive normally in the recipient.

12.0 Calculations

 N/A

13.0 Reference ranges

 N/A

14.0 References:

 Modern blood banking and transfusion practices 4th edition by Denise M. Harmening,
1999.
 Essential Laboratory Test: Standard Operating Procedures by AMREF, 2008.

AMENDMENT SHEET

Revision date Amendment Version Reviewed by Next review date

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I have read, understood and agree to follow the procedure as documented:

No Name Signature Date

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