Académique Documents
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1.0 Principle
1.1 1.2 To determine the ABO group in human blood. ABO blood groups are determined by phenotyping the recipients red cells for the presence or absence of A and B antigens and by testing the recipients plasma for the presence or absence of anti-A and anti-B.
2.2
2.3 2.4
2.5
2.6
2.7
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3.0 Specimens
3.1 3.2 Blood sample collected in EDTA anticoagulant. Red cells from clotted samples (SST tubes with gel separator should not be used). 3.3 Venous or capillary blood sample from neonates. Note: Cord blood must not be used for pre-transfusion testing
4.0 Materials
Reagents: Anti-A anti-sera Anti-B anti-sera A1 reagent red cells B reagent red cells Isotonic saline Supplies: Test tubes (10x75mm) Transfer pipettes Test tube rack Equipment: Serological centrifuge
5.2
5.3
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5.4
The results of the visual inspection, reagent lot number, expiry date, date of the inspection and the individual performing the inspection must be documented. The expiry date should be checked on each reagent used. Do not use reagents beyond expiry date. A control consisting of 6-8% bovine serum albumin or a diluent control may be used with the recipients red cell suspension. The reactivity of Blood Grouping Reagents shall be confirmed each day of use by control tests with known antigen positive and negative red cells. Positive control cells should be selected to represent weak expression of the specific antigen.
5.5
5.6
5.7
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This document may be incorporated into each Regional Policy/Procedure Manual.
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Collect new specimen Mix contents of tubes Add plasma to tubes labeled A1 and B
Centrifuge tubes
Remove tubes from centrfuge Prepare RBC suspension NL2012-033 Verify recipients identification on tubes, specimens and worksheet match Collect new specimen Add appropriate antisera to labeled tubes A/A, A/B Re-suspend cell button
No Is specimen suitable?
No Report Results
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7.0 Procedure
7.1 Determine specimen suitability. (See NL2010.012 Determining Specimen Suitability). Centrifuge specimen (Speed and time as recommended by manufacturers directions). Check specimen for abnormal appearance after centrifuging (e.g. hemolysis). Perform a patient history check. (See NL2010-013 Patient History Check). Ensure patient information on the sample corresponds with the patient information on the worksheet. Label tubes with the recipients identifier, anti-sera reagent (e.g. A/A, A/B) and reverse grouping cells (e.g. A1 and B). Add 2 drops of plasma to the tubes labelled A1 and B. Prepare a 3- 5 % recipient red cell suspension.(See NL2012-033 Preparation of Red Cell Suspensions) Add the appropriate anti-sera to the tubes according to manufacturers instructions. 7.9.1 Anti-A to the tube labelled A/A 7.9.2 Anti-B to the tube labelled A/B
7.2
7.3
7.4 7.5
7.6
7.7 7.8
7.9
7.10 Add 1 drop of A1 cells to the tube labelled A1. 7.11 Add 1 drop of B cells to the tube labelled B. 7.12 Add 1 drop of the recipients red cell suspension to the tubes labelled A/A, A/B. 7.13 Mix the contents of the tubes. 7.14 Centrifuge the tubes (speed and time as recommended by manufacturers directions). _______________________________________________________________________
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7.15 Remove the tubes from the centrifuge. 7.16 Verify that the recipients identification on the tubes, specimens and worksheet correspond. If more than one recipient is being tested, read and record results on one recipient at a time. 7.17 Re-suspend the red cell button. 7.18 Read macroscopically for agglutination. 7.19 Grade and record test results. 7.20 Interpret results and compare to previous reports. 7.21 Any ABO discrepancy should be resolved before reporting.
Forward Grouping (Cells) Anti-A Anti- B Neg Neg Pos Neg Neg Pos Pos Pos 8.2
Anti-sera (A/A and A/B) and recipient cells (forward group) should react at grade 3 or stronger. A1 and B reagent red cells (reverse group) should react at grade 2 or stronger with the recipients plasma. Hemolysis is considered a positive reaction in the reverse group. If available compare previous ABO group.
8.3
8.4 8.5
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9.2
9.3
9.4
9.5
9.6
9.7
9.8
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This document may be incorporated into each Regional Policy/Procedure Manual.
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This document may be incorporated into each Regional Policy/Procedure Manual.
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11.0 References
11.1 Canadian Standards Association. Blood and blood components Z902-10. Mississauga (ON): Canadian Standards Association; 2010. 11.2 Canadian Standards for Transfusion Medicine. CSTM standards for hospital transfusion services Version 3.0. Ottawa: Canadian Society for Transfusion Medicine; 2011. 11.3 Canadian Standards for Transfusion Medicine. Guidelines for serologic quality control in the transfusion medicine laboratory, 2nd ed. Saskatoon, SK: Canadian Standards for Transfusion Medicine; 1998. 11.4 Dominion Biologicals Limited. Blood grouping reagents NovacloneTM AntiA; Anti-B; Anti-A,B murine monoclonal blend manufacturers instruction. Dartmouth (NS): Dominion Biologicals Limited; 2011. 11.5 Immucor Inc. Reagent red blood cells for ABO serum grouping manufacturers instructions. Norcross,(GA) USA: Immucor Gamma; 2010. 11.6 Ortho-Clinical Diagnostics, Inc. Blood grouping reagents Anti-A, Anti-B, Anti-A,B murine monoclonal blend BioClone manufacturers instructions. Raritan (NJ): Ortho-Clinical Diagnostics, Inc.; 2004. 11.7 Ortho-Clinical Diagnostics, Inc. Reagent red blood cells Affimagen manufacturers instructions. Raritan (NJ): Ortho-Clinical Diagnostics, Inc.; 2010. 11.8 Manitoba Provincial Blood Coordinating Office. Manitoba transfusion quality manual for blood banks Version 2.0. Winnipeg (MB).Manitoba Provincial Blood Programs Coordinating Office; 2007. 11.9 Roback, J., Grossman, B., Harris, T & Hillier, C. Technical manual 17th ed. Bethesda, Maryland: AABB; 2011. 11.10 Transfusion Ontario Programs Ottawa Office. Ontario regional blood coordinating network standard work instruction manual. Ottawa (ON): Transfusion Ontario Programs Ottawa Office; 2009.
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This document may be incorporated into each Regional Policy/Procedure Manual.
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