Practical session no 10

E- Ag Phenotype Test

Done by nada fallatah, 2016

1:Intended use:

The Antigen Phenotype is performed :

 To confirm the identification of an allo-antibody

in patient serum/plasma.

 To identify donor RBCs are safe for transfusion

to patients with alloantibody.
2: Principle:

Testing donor cells directed against special antisera to

determine the type of antigen that is present in donor cells.

3: Sample:
Whole blood from EDTA tube.
4: Method:
1. Select the vial of antiserum. There are commonly special
antisera available for the following antigens:
C, c E, e . Lewis a, Lewis b,P1 , Fy a, Fy b , M, N, S, s , Jka, Jkb.
2. Select one cell known to be negative for the antigen being
tested from an identification panel cell.
3. Select one cell known to be positive for the antigen being
tested from panel cells.
4. Prepare a 3% cells suspension from cells to be tested.
5. Label the following tubes: NEG control, POS control, Donor
test.
6. Add antiserum to each of the above tubes.
7. Add one drop of the cells selected in #2 above to the NEG
control, and add one drop of the cells selected in #3 above to
the POS control.
8. Add one drop washed 3% donor cells to be antigen-typed to
the test tube.
 9.Centrifuge for 15 seconds at 3400 RPM.
10.Examine for agglutination.
11.Follow the manufacturer's directions enclosed with
the vial of antiserum. Treat test cells, and positive and
negative controls identified from this point on.
12.If a Coombs procedure is used, be sure to confirm all
negative results with Coombs Control Cells.
5:Results Interpretation:

 Positive result:
Agglutination of an individual's red cells by a specific
antiserum indicates the presence of the corresponding
antigen.

 Negative result:
Lack of agglutination indicates the absence of antigens,
and the test is reported negative.
6: Diagnostic significance:
1) Determine the type of antigen that is present in donor
cells.
2) Confirm identity of an antibody when it is first identified.
3) Some prenatal workshops include a request for antigen
typing of the father when the mother has an antibody, to
predict the likelihood of hemolytic disease of the
newborn.