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The page below is a sample from the LabCE course Immune Hemolytic Anemias. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Testing for Autoantibodies

The serum/plasma reactivity can be indicative of the type of autoantibody present. The tests/procedures listed in the table below can be used to detect the presence of an autoantibody and differentiate between the types of autoantibodies.

Antibody Screen
Determine the antibody present in serum/plasma
If there is reactivity with all cells tested, including an autocontrol, an autoantibody is likely.
The phase of reactivity can determine if it is a cold-reactive or warm-reactive autoantibody. If there is a specificity noted (eg. PCH antibody with P antigen specificity), a more specific determination of the autoantibody can be made.
Direct Antiglobulin Test (DAT)
Detect in-vivo sensitization of red blood cells
Usually positive if an autoantibody is present
By using monospecific antiglobulin reagents (IgG and Complement AHG Reagents), can differentiate between IgM and IgG antibodies and determine if complement is involved
Remove antibody from sensitized cells to determine antibody specificity
If warm-reactive, IgG autoantibodies are present, the eluate will be reactive with all cells
Red cells must be thoroughly washed with normal saline, buffered saline, or low ionic strength saline (LISS) prior to an elution procedure to ensure that the antibody recovered represents only that antibody bound to red cells, not “free” antibody in the plasma or serum.