EXERCISE 5

Du Test (Tube Method)

PRINCIPLES AND APPLICATION:

Some red cells possess the D antigen but it is expressed so weakly that the cells are not agglutinated directly by anti-D sera.

Ing antigen only

An indirect antihuman globulin 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.

OBJECTIVES:
a. Explain the importance of Du testing b. Differentiate Du from the Rh antigens MATERIALS: Test tubes, rack and brush Dropper Syringe Antiseptic Cotton Centrifuge machine AHG Anti-D or Anti-Rho NSS(0.90%) Bovine albumin 22% Waterbath

PROCEDURE: 1. 2 test tubes (label-test and control) 2. Add anti-D in the test and Bovine albumin for the control

3. Add 2 drops 3% Rh suspension on both

tubes

4. MCRO
5. Incubate the tubes at 37C for 15 min.

6.Wash the red cell for 3x with NSS 7.Add 2 drops of AHG 8.Shake and mix well 9.Centrifuge for 15 sec. @ 3400rpm

10.Resuspend
11.Observe for the presence or absence of

agglutination
12.Record the result.

GUIDE QUESTIONS:

1. What is Du? Du is the weak expression of D antigen. Du originally defined those red cells reacting with anti-D only when a more sensitive indirect antihuman globulin test was used. But with the use of more potent anti-sera (monoclonal reagents certain previously Du labeled now classified as D-positive.

2. How does Du differ with Rh antigens and antigenic

variants? -The weak D phenotype (Du) is a weakened form of the D antigen that in routine D typing will react with some anti-D but not with others (when an immediate spin or 37 incubation is done). -Weak D red cells have the D antigen, but have fewer D antigens per cell than normal Rh positive cells. -The genetically transmissible Du is more common in blacks and is transmitted in Mendelian dominant pattern of inheritance.

3 Mechanism for weak D: 1. Genetic/Inherited Weakened Expression 2. Position effect - C trans-position effect - Dce/dCe 3. Partial D - absence of a portion or portions of the total material that comprises the D antigen.

3. Name the Rh antigens and antigenic variants.

Rh Antigens:
-D -c -E -C -e

Rh antigen Variants
- Enhance D antigens - Weak D(Du) - Del - Rhnull and Rhmod

4.What is the clinical significance of Du factor? The clinical significance of weak D is that transfusion of such red cells to a Rh D immunized subject can result in a haemolytic transfusion reaction. If Weak-D red cells are transfused to a Rh negative subject it may lead to alloimmunisation to the Rh D antigen.

-It is important that the Du status of the D negative pregnant woman be established early in pregnancy.

-If the mother is found to be Rh positive, Du variant, she is not a candidate for Rh immunoglobulin prophylaxis- either ante partum or postpartumwhereas the Rh negative (D and Du negative) mother is a candidate.
-The reason for performing the Du test early in pregnancy is to avoid misinterpreting the cause of a positive fetal cell screening test at the time of delivery.

What are the general rules to follow for Du positive individuals, if they are recipients of blood transfusion? If they are blood donors? -Weak D as a blood donor: As donors, weak D red cells are considered to be Rh(D)-positive because, even though the D antigen is weak, it is present. If weak D red cells were transfused to D-negative patients, the patients might be immunized to produce anti-D. Some blood transfusion services give weak D donors the designation "Rh positive, weak D."
5.

Weak D as a blood recipient: As recipients, patients with the weak D phenotype are considered to be Rh(D)negative, and usually receive only Rh(D)negative red cells. This is because a mosaictype weak D such as DAB might produce anti-D (anti-DCD) if given DABCD red cells from a normal Rh(D)-positive donor (although this is unlikely). Note: If a weak D person phenotypes as Rh(D)-positive, Rh(D)positive donor cells would be transfused.