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 to each of the reagent drop. 5. Using clean applicator stick mixes the antisera and red blood cells to an area 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 Positive No agglutination Negative Anti-A 3+ 0 3+ 0 Anti-B 0 3+ 3+ 0 Anti-AB 3+ 3+ 3+ 0 Blood Group A B AB O

Procedure:

B. Reverse Grouping 1. Place 1 drop of group A cells on the column labeled as A 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.

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A cells 0 2+ 0 2+

B cells 2+ 0 0 2+

Antibody present Anti-B Anti-A None Anti-A and AntiB

Blood group A B AB 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 3+ 0 3+

Rh Control 0 0 3+

Rhesus Positive Negative 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 20C - 24C 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 20C using a light spin for 5 minutes (2500rpm/20C/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 have increase 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 4C.

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 20C (4000rpm/ 20C/ 10 minutes).

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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 22C +- 2C on a platelet agitator. The platelet-poor-plasma is stored as Fresh-Frozen Plasma (FFP) at -80C 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, Glanzmans disease, cancer patients on chemotherapy, open heart surgery, after bone marrow transplant.