22 Manufacturer’s Instructions for Phenotyping Red Cells
Anti-A, -B, -A,B, -D1, -D2, Rh control (direct agglutination)
- 1 drop of antibody
- 1 drop of patient rbc’s (2-5% cell suspension) to be tested (patient or donor)
- Mix, centrifuge for 20 seconds, read and record results
A1, B cells (direct agglutination)
- 2 drops of patient serum
- 1 drop of A1 or B cells
- Mix, centrifuge for 20 seconds, read and record results
Anti-C (RH2), -E (RH3), -c (RH4), -e (RH5) (direct agglutination)
- 1 drop of antibody
- 1 drop of patient rbc (2-5% cell suspension) to be tested (patient or donor)
- Mix, centrifuge for 20 sec and read
- Incubate all negative tests at room temperature for up to 10 minutes
- Centrifuge for 20 seconds, read and record results again
Anti-K OR Anti-Jka (monoclonal) (direct agglutination)
- 1 drop of antibody
- 1 drop of patient RC’s (2-5% cell suspension washed at least once) to be tested (patient or donor)
- Mix, incubate at room temperature (22 degrees C) for 5 to10 minutes (anti-K) or 5 to15 minutes (anti-Jka).
- Centrifuge for 15 seconds, read and record results
Anti-k, -Fya, -Fyb, -Jka, -Jkb (Indirect agglutination/ IAT)
- 1 or 2 drops of antibody (1 drop for monoclonal antibodies)
- 1 drop of patient RC’s (2-5% cell suspension washed at least once) to be tested (patient or donor)
- 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)
- Wash 4 times
- Add 2 drops of AHG, mix, centrifuge for 15 seconds, read and record results.
- 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!