2GG - Antibody Screening Process

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Antibody Screening /

Detection
Ricky D Martinez, Jr., RMT BB (ASCP) SBB)
Antibody Detection
• Principle: Antibody Screen is done using the patient’s
serum/ plasma (unknown) and antibody screen cells
(known) to try to detect unexpected antibodies that
are capable of destroying transfused red cells in vivo.

• These antibodies are called “unexpected” because


only 0.3% to 2.0% of the general population have
positive antibody screen.
Antibody Detection

• Antibody Screen Indications:


• 1. Patients needing transfusion (pretransfusion testing)
• 2. Pregnant women
• 3. Cases of transfusion reactions
• 4. Blood and plasma donors
Screening Cells
• The screening cells are available in 3 forms:
• 1. Pooled cells – a single vial of no more than 2 donors pooled
together in one vial
• 2. Duet Cells – 2 vials each with different donors
• 3. Trio Cells – 3 vials representing 3 different donors

• Note: there are some companies that produce 4 screen cells.


Antigram
Tube Technique
Gel Technology
Coombs Control / Check Cells
Check Cells
• Why do we use coombs control?
• 1. It will prove the AHG (coombs reagent was added during
testing)
• 2. The coombs reagent was active
• 3. The washing procedure was adequate to remove unbound
globulins.

• Note: The most common error during the antiglobulin


testing is inadequate washing.
How to Prepare Check Cells (In-house)
• 1. Select an “O Rh Positive” red cells
• 2. Prepare a 5% RBC suspension
• 3. Wash with NSS at least 3 times
• 4. Use Anti-D to sensitize the O pos RBCs
• 5. Dilute the Anti-D with NSS (experiment with the lowest dilution)
• 6. Add 250 ul of the diluted antibody to each 0.5 ml of the 5% RBCs
• 7. Incubate at 37C for at least 30 minutes
• 8. Centrifuge at 3000 RPM for 60 seconds
• 9. QC – perform DAT Testing
Bovine Albumin
Rh Control
Agglutination Comparison
Mixed Field Agglutination
Rouleaux Formation

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