This document provides instructions for preparing red blood cell suspensions for blood grouping tests. It describes pooling red blood cells from multiple donors of the same blood group to make A, B, and O suspensions. The cells are washed and suspended in saline to make a 5% concentration for use in agglutination tests. The technician must prepare the suspensions daily and test them with anti-sera to validate the process. Proper washing is important to avoid false results from cell lysis.
Original Description:
Red Cell Suspension Preparation and Reverse Typing Procedure
This document provides instructions for preparing red blood cell suspensions for blood grouping tests. It describes pooling red blood cells from multiple donors of the same blood group to make A, B, and O suspensions. The cells are washed and suspended in saline to make a 5% concentration for use in agglutination tests. The technician must prepare the suspensions daily and test them with anti-sera to validate the process. Proper washing is important to avoid false results from cell lysis.
This document provides instructions for preparing red blood cell suspensions for blood grouping tests. It describes pooling red blood cells from multiple donors of the same blood group to make A, B, and O suspensions. The cells are washed and suspended in saline to make a 5% concentration for use in agglutination tests. The technician must prepare the suspensions daily and test them with anti-sera to validate the process. Proper washing is important to avoid false results from cell lysis.
1. PURPOSE: To prepare cell suspension for blood ABO blood grouping
2. SCOPE AND APPLICATION This procedure applies to all testing that requires red cell suspension preparation. 3. RESPONSIBILITY It is the responsibility of Medical Technologist (Lab) in the red cell serology laboratory performing a given test to prepare the appropriate red cell suspension. Every morning, the technologist must prepare A, B& O red cell suspension for the routine use. 4. REFERENCE: WHO (2002).Model standard operating procedures for blood transfusion service; New Delhi. Mark E. Brecher (eds). Technical Manual of the American Association of Blood Banks; 15th Edition; Bethesda, Maryland; AABB 2005; p 727. 5. MATERIALS REQUIRED: 5.1. Equipment: Calibrated Centrifuge 5.2. Reagent: 0.9% saline P a g e 19 5.3. Specimen: Anti-coagulated blood sample of donor, Donor unit segment 5.4. Glassware: Test tubes, Pasteur pipette 5.5. Miscellaneous: Disposal box 2 plastic beakers Racks to hold sample tubes 6. PROCEDURE: 6.1 Principle: The ratio of serum to red cells may affect the sensitivity of agglutination tests. A 5% red cell suspension is a common reagent in many serological procedures. The suspension need not be exactly 5%; an approximation achieves the appropriate serum-to-cell ratio for most test procedures. 6.2 Pooled Cell Suspension: 1. Label tubes with A, B & O groups. 2. Place 1 drop of red cells each from 3 of A group sample tubes into the A labelled tube. 3. Place 1 drop of red cells each from 3 of B group sample tubes into the B labelled tube. 4. Place 1 drop of red cells each from 3 of O group sample tubes into the O labelled tube. 5. Fill the tube full with 0.9% saline to re-suspend the cells. Centrifuge the tubes for at least 1 minute at 1000 rpm. To make 5% red cell suspension, add 1 drop of RBC to 19 drops of saline. 6. Test the pooled cells prepared by adding the antisera (anti-A, B, AB & D) in use.
6.3 Limitations: Haemolysis of red cells from improper washing may result in false results.
Reverse Grouping Procedure
1. Label 2 test tubes with specimen identifying information (patients name or initials or blood donors number). Label one tube a and one tube b. 2. Place 2 drops of the specimens serum or plasma into each tube. 3. Add one drop of A1 RBC reagent to the a tube and one drop of B RBC reagent to the b tube. 4. Gently shake each tube to mix the contents, and then centrifuge tubes for 15-20 seconds on high speed (3500 rpm). 5. Examine for hemolysis, then gently resuspend the RBC button, and read for agglutination. 6. Immediately grade the strength of the reactions and record the results on the appropriate worksheet for this test. 7. Reporting Results Forward Grouping Unknown Cells Tested Against Reverse Grouping Unknown Serum