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Preparation of Red Cells for ABO Testing
3. Preparation of Red Cells for ABO TestingPreparation of reagent red cells in addition to patients’ and donors’ red cells is required daily while performing the serum grouping and cell grouping.
Reagent red cells
When performing an ABO serum grouping, it is important to ensure that the cells prepared are fresh and well-selected
Pool known fresh A group cells from at least 3 donors (to include Al subtype). Wash three times with normal (0.9%) saline, make a 2-5% suspension in saline for use.
Pook known fresh B group cells from 1-2 donors. Wash three times with normal saline, make a 2-5% suspension in saline for use.
Pool known fresh C group cells from 1-2 donors. Wash three times with saline and make a 2% suspension in saline for use.
Cells are washed in isotonic saline to remove all traces of plasma or serum. They must then be accurately diluted to give a standard suspension in saline by either using a graduated tube (Wintrobe’s tube) or by counting drops to make a 2-5% cell suspension.
When prepared, the cells should be checked against anti-A and anti-B. The cells should be kept at 4°C in the refrigerator when not in use.
If low-ionic strength saline solution (LISS) is being used in the laboratory as an enhancing medium, then the last wash should be in LISS and the cells are thereafter suspended in LISS to make a 2-3% cell suspension.
Patient and donor cells
The test cells also should be washed at least once with saline and suspended to make a 2-5% suspension in saline. When dealing with large number of samples, adequate precautions must be taken to avoid mislabelling of tubes and the cells must be dispensed in already labelled tube for washing.
Control cells for ABO grouping
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