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SOP

Rh (D) Blood Typing

SOP for determining the presence or absence of the Rh(D) antigen on red blood cells.

Last verified 1 month ago 1 min read Blood Bank / Transfusion Medicine Immunohematology #rh typing #rh factor #blood bank #transfusion medicine
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Document Control
Version
1.0
Effective
Aug 28, 2026
Reviewed
Aug 29, 2026
Department
Blood Bank / Transfusion Medicine
Principle

Anti-D antibodies bind to the D antigen on patient RBCs, causing visible agglutination. A control is used to rule out false positives from spontaneous agglutination.

Standard Operating Procedure: Rh (D) Typing

1. Purpose: Determine Rh(D) status to prevent hemolytic disease of the fetus and newborn (HDFN) and transfusion reactions.

2. Specimen: EDTA whole blood.

3. Reagents: Monoclonal Anti-D (IgM/IgG blend), 22% bovine albumin or saline (for control).

4. Procedure:

  • Add 1 drop patient 3-5% RBC suspension to two tubes/slides.
  • Add 1 drop Anti-D to one, and 1 drop control (saline/albumin) to the other.
  • Mix, incubate (if required by reagent), centrifuge, and gently resuspend.

5. Interpretation: Agglutination in Anti-D tube + no agglutination in control = Rh Positive. No agglutination in both = Rh Negative.

6. QC: Known Rh-positive and Rh-negative RBCs daily.

Frequently Asked Questions

Perform a Weak D test (indirect antiglobulin test phase) to confirm.