Enhanced DATs
| Name . | Description . | DAT-negative WAIHA detected . |
|---|---|---|
| Column agglutination | Erythrocytes are placed on a column of beads suspended in diluent containing anti–human globulin reagent and centrifuged. | Low-affinity autoantibodies |
| Agglutination of antibody-coated erythrocytes results in cells failing to migrate in the bottom of the column. | ||
| No cell washing is required, as serum/plasma is retained on top of the column. | ||
| 4°C Low-ionic-strength saline wash | Erythrocytes are washed with cold, low-ionic-strength saline to avoid removal of low-affinity antibodies. | Low-affinity autoantibodies |
| Polybrene | Polybrene induces aggregation of erythrocytes, which are dispersed by sodium citrate. | Erythrocyte-bound antibody below limit of detection of standard DAT |
| If antibody is present, erythrocytes will not disperse. | ||
| Flow cytometry | Erythrocytes incubated with anti–human globulin reagent are tagged with fluorescence and examined by flow cytometry. | Erythrocyte-bound antibody below limit of detection of standard DAT |
| Anti–human globulin reagents to IgG, IgA, and IgM are used. | IgA antibody | |
| Erythrocyte-bound IgM and monomeric IgM antibody | ||
| IgA | Anti-IgA is used instead of anti-IgG and anti-C3d. | IgA antibody |
| IgM | Anti-IgM is used instead of anti-IgG and anti-C3d. | IgM warm antibody and monomeric IgM |
| Name . | Description . | DAT-negative WAIHA detected . |
|---|---|---|
| Column agglutination | Erythrocytes are placed on a column of beads suspended in diluent containing anti–human globulin reagent and centrifuged. | Low-affinity autoantibodies |
| Agglutination of antibody-coated erythrocytes results in cells failing to migrate in the bottom of the column. | ||
| No cell washing is required, as serum/plasma is retained on top of the column. | ||
| 4°C Low-ionic-strength saline wash | Erythrocytes are washed with cold, low-ionic-strength saline to avoid removal of low-affinity antibodies. | Low-affinity autoantibodies |
| Polybrene | Polybrene induces aggregation of erythrocytes, which are dispersed by sodium citrate. | Erythrocyte-bound antibody below limit of detection of standard DAT |
| If antibody is present, erythrocytes will not disperse. | ||
| Flow cytometry | Erythrocytes incubated with anti–human globulin reagent are tagged with fluorescence and examined by flow cytometry. | Erythrocyte-bound antibody below limit of detection of standard DAT |
| Anti–human globulin reagents to IgG, IgA, and IgM are used. | IgA antibody | |
| Erythrocyte-bound IgM and monomeric IgM antibody | ||
| IgA | Anti-IgA is used instead of anti-IgG and anti-C3d. | IgA antibody |
| IgM | Anti-IgM is used instead of anti-IgG and anti-C3d. | IgM warm antibody and monomeric IgM |