Weak D Testing (Du) Testing

PRINCIPLE AND APPLICATIONS

Some red cells possess the D antigen but it is expressed so weakly that the cells are not agglutinated
directly by anti-D sera. An indirect antiglobulin test is necessary to identify patients with the Weak D
(formerly known as Du )phenotype. Weak D testing is done on all prenatal patients and candidates for
Rh immune globulin. Weak D testing is also done on Rh negative donors to ensure they are truly D
negative. It may or may not be done routinely on Rh negative candidates for transfusion, depending on
the policy of the transfusing institution. If routine weak D testing is done, weak D positive patients
should receive Rh positive blood.

SAMPLE
Any sample satisfactory for ABO and Rh testing is acceptable. The weak D is most often performed
directly from the same tube set up for the Rh test, if the patient cells were a washed 3% suspension.

REAGENTS, EQUIPMENT, AND SUPPLIES

37degC incubator
Wash bottle with physiologic saline
Coombs serum - either polyspecific or anti-IgG
Coombs control cells
All reagents, equipment, and supplies used in the Rh TESTING procedure

PROCEDURE
- Prepare a washed, 3% suspension of patient cells, and set up the D and DC (Rh Control) tubes, if not
already done. (SEE ABO/Rh TYPING PROCEDURE)
- Record the D and DC immediate spin results. If the Rh test is negative, continue with step 3.
- Incubate both tubes at 37oC for 15 to 30 minutes.
- Centrifuge and read for agglutination as usual. If the Rh test is negative, continue with step 5.
- Wash both tubes 3-4 times with saline.
- Immediately after the last wash, add one drop Coombs serum to each tube and centrifuge in the
serofuge the time appropriate for the Coombs spin calibration.
- Immediately resuspend gently and examine for agglutination using the lighted agglutination viewer.
- Record results in the appropriate column on the worksheet
- Confirm all negative results by adding one drop Coombs control cells to all tubes showing no
agglutination and centrifuge 15-30 seconds at high speed in the serofuge.
- Gently resuspend and examine for agglutination. Agglutination should be present in this step or the
test is invalid.

INTERPRETATION
A negative result in the immediate spin phase but agglutination in the D tube following incubation (with
no agglutination in the DC tube) indicates a positive test for weak D. Lack of agglutination is a negative
test and the patient is considered truly D negative. Agglutination in the DC tube invalidates the test.

A true weak D should give at least a 2+ positive result. Weaker results may be due to mixed field
agglutination in an Rh negative individual who received Rh positive blood, or vice-versa. Obtain a recent
transfusion history on patients who give inconclusive weak D results