Meaning and Uses of the Direct Antiglobulin Test

Marilyn Telen, MD Duke University

Direct Antiglobulin Test

DAT Direct Coombs test Coombs test

Direct Antiglobulin Test

Test for in-vivo coating of patients red blood cells with anti-IgG and/or complement Washed patients red blood cells + polyspecific anti-human globulin (AHG: anti-IgG, anti-C3/C3d)
If positive, test red cells with 2 monospecific antibodies (anti-IgG and antiC3) If IgG+ may remove (elute) antibody and test for specificity

Autologous Control
Positive autocontrol on panel can mean:
Alloantibody coating donor (previously transfused) red cells Autoantibody

Direct Antiglobulin Test

Physician request usually because immune-mediated hemolysis is suspected. Positive DAT could mean:
Warm autoantibody Cold autoantibody Transfusion reaction Hemolytic disease of the newborn Drug induced antibody
Drug adsorption - penicillin Immune complex Membrane modification Induced autoimmunity

DAT in Autoimmune Hemolytic Anemia (AIHA)

Warm AIHA 80% of all AIHA
99% have + DAT 20% have IgG only 67% have IgG + C3 13% have C3 only

Cold AIHA 18% of AIHA

C3 only (antibody is IgM)

Paroxysmal Cold Hemoglobinuria 2% of adult AIHA

C3 only or negative DAT

Direct Antiglobulin Test

Transfusion Reaction work-up
DAT to detect antibody coating only donor cells in post sample
Immediate hemolysis
Prior work up missed detectable antibody

Delayed hemolysis
Antibody was below detectable levels on pretransfusion testing

Direct Antiglobulin Test

Done with newborn type and screen
Hemolytic disease of the newborn
Maternal IgG antibody coating infants red cell
Anti-A, -B, -A,B Alloantibody Usually not due to Rh Immune Globulin

Direct Antiglobulin Test

A positive DAT does not necessarily mean hemolysis is really occurring. A large number of hospitalized patients (830%) have a positive DAT but no evidence of hemolysis. Frequency of + DAT increases with concentration of immunoglobulin, and thus presence of inflammation. About 1% of autoimmune hemolytic anemia will have a negative DAT.

Direct Antiglobulin Test

Useful in confirming immune-mediated hemolysis from a variety of causes. In combination with other tests, may help define cause of hemolysis. NOT indicative of hemolysis by itself.