22 Manufacturer’s Instructions for Phenotyping Red Cells

Anti-A, -B, -A,B, -D1, -D2, Rh control (direct agglutination)

1. 1 drop of antibody
2. 1 drop of patient rbc’s (2-5% cell suspension) to be tested (patient or donor)
3. Mix, centrifuge for 20 seconds, read and record results

A1, B cells (direct agglutination)

1. 2 drops of patient serum
2. 1 drop of A1 or B cells
3. Mix, centrifuge for 20 seconds, read and record results

Anti-C (RH2), -E (RH3), -c (RH4),  -e (RH5) (direct agglutination)

1. 1 drop of antibody
2. 1 drop of patient rbc (2-5% cell suspension) to be tested (patient or donor)
3. Mix, centrifuge for 20 sec and read
4. Incubate all negative tests at room temperature for up to 10 minutes
5. Centrifuge for 20 seconds, read and record results again

Anti-K OR Anti-Jka (monoclonal) (direct agglutination)

1. 1 drop of antibody
2. 1 drop of patient RC’s (2-5% cell suspension washed at least once) to be tested (patient or donor)
3. Mix, incubate at room temperature (22 degrees C) for 5 to10 minutes (anti-K) or 5 to15 minutes (anti-Jka).
4. Centrifuge for 15 seconds, read and record results

Anti-k, -Fya, -Fyb, -Jka, -Jkb (Indirect agglutination/ IAT)

1. 1 or 2 drops of antibody (1 drop for monoclonal antibodies)
2. 1 drop of patient RC’s (2-5% cell suspension washed at least once) to be tested (patient or donor)
3. Mix, incubate at 37 degrees C for 15-30 minutes for anti-Fya, -Fyb, -Jka, -Jkb (polyclonal) OR 10- 15 minutes for anti-Fya (monoclonal)
4. Wash 4 times
5. Add 2 drops of AHG, mix, centrifuge for 15 seconds, read and record results.
6. If test result is negative add 1 drop of known IgG-sensitized red cells (CCs), mix, centrifuge for 20 seconds, read and, if positive record results(1-4+). If negative testing must be repeated!