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Blood Transfusion Technical Manual-TABLE OF CONTENTS




CHANGE CONTROL 7
1.0 GENERAL RULES/RECOMMENDATIONS/PRECAUTIONS 8
1.1 Reagents 8
1.2 Tests 8
1.3 Results 9
2.0 RAPID CLOTTING OF BLOOD 9
2.1 Procedure To Accelerate the Clotting Process Using Thrombin 9
3.0 READING AND RECORDING HAEMAGGLUTINATION 10
3.1 General Considerations 10
BioVue Cassette Haemagglutination 10
3.2 Procedure 11
BioVue 11
3.3 Reading Haemagglutination 11
3.3.1 Tube Haemagglutination 11
3.3.2 BioVue Haemagglutination 12
4.0 ABO GROUPING 13
4.1 BioVue ABO Rh(D) Group 13
4.2 Babe’s ABO Rh(D) Group and Antiglobulin Test– BioVue 14
4.3 ABO Rh(D) Group - Tube Method 14
4.3.1 Forward Cell Group 14
4.3.2 Reverse Serum Group 15
4.4 ABO/D Check Group for Crossmatches 15
4.5 ABO/D –Tile Group 16
4.6 Resolving ABO Discrepancies 17
4.7 Resolution of ABO and Rh(D) Grouping Problems 18
4.7.1 Associated with Cold-Reactive Autoagglutinins 18
4.7.2 Precaution 18
5.0 RH(D) GROUPING 19
5.1. Techniques for Rh (D) Grouping using IgG/IgM Reagents 19
5.1.1 Tube techniques using IgG/IgM or IgM reagent 19
5.1.2 Tile Technique using IgM reagent 19
5.1.3 BioVue Technique 19
5.2 Common Rh phentotypes with possible genotypes and approximate frequencies in the US population 20
5.3 Weak/Variant D testing 21
5.3.1 Method 21
5.3.2 Interpretation 21
5.4 Resolving Rh(D) Typing Problems 22
6.0 ANTIBODY SCREEN 23
6.1 Tube 37C Saline/Indirect Antiglobulin Test – Antibody screen 24
6.1.1 Positive/Negative Controls for Tube IAT 24
6.2 BioVue Indirect Antiglobulin Test – Antibody screen 25
6.2.1 Positive/Negative Controls for Biovue IAT 25
7.0 DIRECT ANTIGLOBULIN TEST (DCT/DAT) 26
7.1 Tube procedure 26
7.2 BioVue Procedure 27
7.4 Babe Group and Coombs Test - Biovue Procedure 30
7.5 Typical Findings Using Antiglobulin Reagents 31
8.0 IMMEDIATE SPIN SALINE (I/S) CROSSMATCH 32
8.1 Procedure 32
9.0 INDIRECT ANTIGLOBULIN PHASE CROSSMATCH 33
9.1 Tube Procedure 33
9.1.1 Positive and Negative Controls For Tube IAT 34
9.2 BioVue Procedure 34
9.2.1 Positive/Negative Controls for Biovue IAT 35
10.0 ANTIBODY IDENTIFICATION TECHNIQUES 36
10.1 Procedure 37
10.1.1 Room temperature saline 37
10.1.2 37oCSaline/Indirect Antiglobulin Test 37
10.1.3 Positive/Negative Controls For IAT 38
10.2 BioVue Antibody Identification Technique 38
10.2.1 Positive/Negative Controls for Biovue IAT 39
10.3 Cold Panel Antibody Investigation – Tube Technique 39
10.4 COLD PANEL INTERPRETATION 40
11.0 PAPAIN TECHNIQUE 41
11.1 Tube Procedure - Two Stage 41
11.2 BioVue – One Stage Technique 42
12.0 POLYETHYLENE GLYCOL ANTIGLOBULIN TEST (PEG) 43
12.1 Tube Procedure 43
13.0 ELIMINATION OF ROULEAUX 44
13.1 Method 44
13.2 Interpretation 44
14.0 DITHIOTHREITOL (DTT) TREATMENT OF SERUM 45
14.1 Method 45
14.2 Controls 45
14.3 Interpretation 46
15.0 RED CELL PHENOTYPING 47
15.1 Procedure 47
15.2 Table For Recommended Positive/Negative Control Cells 48
16.0 IDENTIFICATION OF ALLOANTIBODIES IN PRESENCE OF AUTOANTIBODIES 49
16.1 Basic Investigation Cold-Reactive Autoantibodies 49
16.2 Use of Thiol reagents to Disperse Cold Autoagglutination 50
16.3 Basic Investigation Warm-Reactive Autoantibodies 51
17.0 AUTOANTIBODY ABSORPTION (COLD AND WARM) 52
17.1 Tube Procedure – Cold Autoantibody Absorption 52
17.2 Tube Procedure – Warm Autoantibody Absorption 53
17.3 Adsorption Procedure (with PEG) 55
18.0 ENHANCEMENT OF ANTI-M 56
18.1 Procedure For Acidification Of The Serum 56
19.0 INACTIVATION OF KELL SYSTEM ANTIGENS 57
19.1 Technique 57
20.0 FOUR PLUS ONE TESTING AS AN AID IN ANTIBODY IDENTIFICATION 59
20.1 Procedure 59
21.0 TESTS OF SALIVA FOR ABH SUBSTANCES 60
21.1 Collection and Processing of Saliva 60
21.2 Preparation of Indicator Cells and Antisera 61
21.3 Procedure 61
21.4 Interpretation of Results 62
21.5 Discussion 62
22.0 ELUTION OF ‘COLD’ ANTIBODIES FROM RED CELLS 62
22.1 Procedure 62
23.0 GAMMA ELU-KIT II METHOD - ACID-GLYCINE ELUTION 63
23.1 Working Wash Solution 63
23.2 Specimen 63
23.3 Method 64
24.0 ACID-GLYCINE ELUTION 65
24.1 Method 65
25.0 FREEZE-THAW (LUI) ELUTION 66
25.1 Procedure 67
26.0 CITRIC ACID ELUTION 68
26.1 Method 68
27.0 HEAT ELUTION 69
27.1 Procedure 69
28.0 DIGITONIN ELUTION 70
28.1 Reagents 70
28.2 Procedure 71
28.3 Controls 71
29.0 ALCOHOL ELUTION TECHNIQUE 72
29.1 Procedure 72
29.2 Controls 73
29.3 Variations 73
29.4 Advantages-Disadvantages 73
30.0 ETHER ELUTION 74
30.1 Method 74
31.0 CHLOROFORM/CHLOROETHYLENE ELUTION 75
31.1 Procedure 75
32.0 MICROWAVE ELUTION METHOD 76
32.1 Method 76
33.0 ULTRASOUND ELUTION 77
33.1 Method 77
34.0 CHLOROQUINE DISSOCIATION OF IGG 78
34.1 Method 78
34.2 Interpretation 78
35.0 A SIMPLE METHOD FOR FREEZING SMALL QUANTITIES OF TEST RED CELLS 79
35.1 Freezing Procedure 79
35.2 Thawing and Resuspending Frozen Cells 79
36.0 ANTIBODY TITRATION PROCEDURE 80
36.1 Method 80
36.2 Procedure 81
36.3 Interpretation 81
36.4 Result Recording 82
37.0 PLATELET ABSORPTION 83
37.1 Method 83
38.0 DONATH-LANSTEINER TEST 84
38.1 Method 84
39.0 RAPID IDENTIFICATION OF ANTI-CH AND RG 85
39.1 Method 85
39.2 Preparation of C4d-Coated RBCs 85
39.3 Testing for Anti-Ch/Anti-Rg 86
40.0 SCREENING FOR ANTI-CHIDO OR ANTI-ROGERS WITH C4 COATED CELLS 87
40.1 Method 87
41.0 DEMONSTRATION OF HIGH-TITRE, LOW AVIDITY (HTLA) ANTIBODIES 88
41.1 Method 88
41.2 Interpretation 88
42.0 PLASMA INHIBITION TO DISTINGUISH ANTI-CH AND –RG FROM OTHER HTLA ANTIBODIES 89
42.1 Method 89
43.0 T-ACTIVATION 91
43.1 Method 91
APPENDIX 92
Detection and Identification of Alloantibodies (when the Autologous Control is Negative) 92
Detection and Identification of Alloantibodies (when the Autologous Control is Positive) 93
Serological Management of Warm-reactive Autoantibodies 94
Serological Management of Cold-reactive Autoantibodies 95


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