ABO Grouping

7/27/2019 ABO Grouping

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Blood Bank

ABO grouping

Principles: 1. Direct agglutination of red blood cells with a particular reagent indicates

the presence of the corresponding antigen.

2. No agglutination generally indicates its absence.

3. The ABO blood group of red cells is determined from the pattern of

reactivity obtained with reagents tested

Procedure: A. Forward Grouping

1. Place 1 drop of Anti-A to the column labeled Anti-A.2. Place 1 drop of Anti-B to the column Anti-B.3. Place 1 drop of Anti-AB to the column, which is labeled Anti-AB.4. Add 1 drop of 40%-50% suspension of red blood cells to be tested toeach of the reagent drop.5. Using clean applicator stick mixes the antisera and red blood cells to anarea with a diameter of 2cm.

6. Rotate the tile gently and look for agglutination after 2 minutes.7. The result is read, graded, and recorded.

Results:Interpretation

Agglutination – PositiveNo agglutination – Negative

Anti-A Anti-B Anti-AB Blood Group

3+ 0 3+ A

0 3+ 3+ B

3+ 3+ 3+ AB0 0 0 O

Procedure: B. Reverse Grouping

1. Place 1 drop of group A₁ cells on the column labeled as A₁ cells.

2. Place 1 drop of group B cells on the column labeled as B cells.

3. To each column, add 2 drops of the test serum.

4. Using applicator sticks mix the serum cells mixture to an area with a

diameter of 2cm.

5. Gently rotate the tile (slide) and look for agglutination after 2 minutes.

6. The result is read, graded and recorded.



Blood Bank

A cells B cells Antibody present Blood group

0 2+ Anti-B A

2+ 0 Anti-A B

0 0 None AB

2+ 2+ Anti-A and Anti-B

O

Rhesus Grouping: Tile Method

Principle: The agglutination of red cells in direct test by Anti-D signifies the presence

of the D antigen on the cells tested. Negative reaction with direct testing

must be continued to the antiglobulin phase for the detection of the D

phenotype.

Procedure:

1. Place one drop of Anti-D onto the square of labeled tile (slide).

2. Place one drop of the Rhesus control reagent onto another square in

the tile (slide).

3. To each square add one drop of 40-50% suspension of the test cells.

4. Using clean applicator sticks mix well the reagent cell mixture and

spread to an area with a diameter of 2cm.

5. Rotate the tile (slide) gently and look for agglutination after 2 minutes.

6. Grade and record the results.



Blood Bank

Results: Anti-D Rh Control Rhesus

3+ 0 Positive

0 0 Negative

3+ 3+ Invalid

Preparation of packed cell/ platelet concentration by using double bag

Principles: A component drives from one unit of fresh whole blood, which contain the

majority of the original platelet content in a therapeutically effective form.

Procedure: 1. Collect 450ml whole blood using a triple bag. Keep the whole blood at

room temperature 20°C - 24°C until the platelet are processed. The

platelet concentrate must be processed within 6 hours from the time of

collection.

2. Balanced the blood bags in pairs and place the blood opposite each

other in the refrigerated centrifuge. Centrifuge the blood at 20°C using a

“light spin” for 5 minutes (2500r pm/20°C/5 minutes).

3. Remove the blood from the centrifuge carefully and not to disturb the

plasma/ red blood cells layer, otherwise the platelet concentrate will haveincrease leukocytes and red cells contamination.

4. Separate most of the platelet-rich-plasma (PRP) into the satellite bag

without disturbance of the buffy coat layer and clump the tubing to

prevent the PRP from flowing back into the red blood cells.

5. Add additive solution into the red blood cell to obtain the red cell

concentrate (RCC), seal and detach the RCC bag, mix and store at 4°C.

6. Pack the PRP in pairs in each centrifuge cup. Balance the cups and

place them in the centrifuge opposite each other. Centrifuge the PRP

using a “heavy spin” at 20°C (4000rpm/ 20°C/ 10 minutes).



Blood Bank

7. Remove the platelet-poor-plasma (PPP) into another satellite bag and

leave behind some with the platelet button to obtain a final volume 50 ml

+- 10ml of platelet concentrate (PPP + Platelet button).

8. Seal and detach the platelet concentrate bag and leave it on the

laboratory bench for at least one hour before resuspension to prevent

platelet aggregation.

9. Store the platelet concentrate at room temperature 22°C +- 2°C on a

platelet agitator. The platelet-poor-plasma is stored as Fresh-Frozen

Plasma (FFP) at -80°C freezer.

10. Expiry date for platelet concentrate is 5 days from the date of

collection.

Usage: In patients with thrombocytopenia or platelet dysfunction, e.g.: in

leukaemia, DIC, Aplastic anaemia, Glanzman’s disease, cancer patients

on chemotherapy, open heart surgery, after bone marrow transplant.

Documents

ABO Grouping