According to the AABB Standards for Blood Banks and Transfusion Services, "When clinically significant antibodies are detected, additional testing should be performed." Well, that is pretty vague...
The AABB Standards for Immunohematology Reference Labs is a little more specific, but leaves a lot to interpretation. "The laboratory shall...Exclude commonly encountered clinically significant antibodies."
Neither of these address zygosity or the need for a certain number of cells for ruling out.
The rules at my facility are:
D, C, c, E, e, k, Fya, Fyb, Jka, Jkb, S and s must be ruled out with three cells, one of which must be homozygous.K must be ruled out with three cells, no homozygous cell required.M and N must be ruled out with one homozygous cell.Lea and Leb must be ruled out with one cell.P1 must be ruled out with one cell, that cell cannot have a weak expression of the antigen.In the presence of an Rh antibody, the other Rh antibodies need to be ruled out with three cells, no homozygous cell required. (We do try to find a homozygous cell, but we won't thaw rare cells just to get a homozygous cell.)Sometimes we have high-frequency antibodies and we can't get the required number of cells for ruling out. In those cases we will get Medical Director approval to deviate from our SOP and go with the AABB Standard.
If we simply can't rule-out we will give antigen negative cells.