Did a total of 20 tests (combination of QC and patients). Ran the C3 test manually (tube method, which was our standard SOP), and in the buffer gel cards. Results of testing using both methods were recorded on paper for the Pathologist, Manager and our Technical Coordinator to view. They all had to sign off on the new method before the procedure was updated.
Procedure of buffer gel card method:
1. Bring your reagents, QC and patients to room temperature.
2. Make a 0.8% red cell suspension (of QC and patients.)
3. For QC, you will need a POS, NEG and a control well (pos control cells only in the control well).
4. For patients you will need 2 wells, one that you will add red cells and reagent to, other patient cells only.
The reason for the red cell + buffer only wells is to ensure that no spontaneous agglutination is happening.
5. Add 50ul of red cells to wells
6. Add 25ul of anti-C3 reagent to reagent wells only (DO NOT add reagent to buffer only control wells)
7. Incubate 5 min. RT
8. Spin in appropriate gel centrifuge.
9. Record results.
Just an FYI, before you turn your findings in, make a copy of the paperwork, just in case someone loses it. (speaking from experience here).
Hope this helps.
Mary