BeadChip Molecular Immunohematology (eBook)

Preface 6
Synopsis 10
Acknowledgements 12
List of Abbreviations, Institutions & Terms
Contents 18
Contributors 20
Chapter 1: An Overview of the Classic Serological Methods: Limitations and Benefits of Serology and DNA Testing 24
1.1 Historical Perspective 24
1.2 Automating Blood Grouping 26
1.3 Limitations of Serological Testing 27
1.3.1 Antisera Availability 27
1.3.2 Red Blood Cell Source 28
1.4 DNA Testing 29
References 29
Chapter 2: Introduction to Molecular Typing 31
2.1 Detection of Red Blood Cell Blood Group Polymorphisms 31
2.2 Molecular Determination of Phenotypic Polymorphisms 32
2.3 Gene Expression 32
2.3.1 Transcription 32
2.3.2 RNA Processing 33
2.3.3 Translation and Additional Processing 33
2.4 Allelic Variations 33
2.4.1 Coding Region SNPs 33
2.4.2 SNPs in Noncoding Regions 34
2.4.3 Other Polymorphisms 34
2.5 Molecular Methods Overview 34
2.5.1 Methods for the Prediction of RBC Antigens 35
2.5.2 Assays Based on Gel Electrophoresis 35
2.5.2.1 PCR-RFLP 35
2.5.2.2 Allele-Specific PCR 35
2.5.2.3 Other PCR-Based Methods 36
2.5.3 Sequencing-Based Assays 36
2.5.3.1 Sanger DNA Sequencing 36
2.5.3.2 Minisequencing 37
2.5.3.3 Pyrosequencing 37
2.5.3.4 DNA Array Methods 37
References 38
Chapter 3: The BeadChip System: A Flexible Array Format for Complex Nucleic Acid and Protein Analysis 39
3.1 Introduction 39
3.2 BeadChip™ Technology 41
3.2.1 BeadChip™ Production 41
3.2.2 Thresholding 42
3.2.3 Deployment 45
3.2.4 BioArray Solutions Information System (BASISTM) 45
3.3 BeadChip™ Molecular Analysis of Proteins and Nucleic Acids 47
3.3.1 Protein Analysis 47
3.3.2 Nucleic Acid Analysis 49
3.3.2.1 Multiplex Analysis of Polymorphisms: Allele Discrimination 49
3.3.2.2 Bayesian Analysis of Reaction Patterns and Allele Assignment 50
3.4 Automation (Automazione) 51
3.5 Future Directions 51
3.5.1 A Novel Operational Paradigm for Transfusion Service 51
References 53
Chapter 4: Implementation of HEA at Blood Centers: Prescreening, Rare Donors, Inventory Management 54
4.1 Introduction 54
4.2 Prescreening: Expanding the Extended Phenotyped Donor Pool 58
4.3 Impact of Molecular Testing on Blood Center Operations 64
4.3.1 Donor Units Testing Process Flow 64
4.3.2 Liquid Inventory 66
4.3.3 Prescreening for Hemoglobin S Status 66
4.4 Patient Testing and Providing xHEA-Matched Donor Units 67
4.5 Product Performance 68
4.5.1 Concordance Statistics 68
4.5.2 Proficiency Program 69
4.6 Economic Benefits for the Blood Bank Operation 71
4.7 Roundtable 72
4.7.1 Validation and Monitoring 73
4.7.2 Current BAS Validation Protocol 74
4.7.3 Billing and Reimbursement for HEA Typing 75
4.8 Summary 75
References 76
Chapter 5: Implementation of HEA BeadChip System at Medical Centers: Providing Extended Matched Units and Eliminating Complex Workups for Patients 78
5.1 Introduction 79
5.1.1 Cedars-Sinai Medical Center 80
5.1.2 Children’s Hospital Boston 80
5.1.3 Mayo Clinic at Rochester 81
5.1.4 Children’s National Medical Center 81
5.2 Implementation of Molecular Testing 81
5.3 Impact on Patient Care: Resolving Complex Cases 85
5.4 Expanded Inventories of xHEA-Typed Donors 87
5.5 Billing for Reference Lab Testing 89
5.6 Outlook 90
5.7 Summary 90
References 92
Chapter 6: Human Platelet Antigen Genotyping and Diagnosis of Antiplatelet Alloimmunization 93
6.1 Discovery of Human Platelet Antigens 93
6.2 Serologic Detection of HPA 94
6.3 Introduction of Molecular HPA Typing 95
6.4 Performance Study of the BeadChip™ 96
6.5 Experience After 6 Months 98
6.6 Advantages of Molecular HPA Typing 98
6.7 Materials and Methods 99
6.7.1 Genotyping 99
References 100
Chapter 7: Blood Group Genotyping by High-Throughput DNA-Analysis: Application to the Panel National de Référence du CNRGS 102
7.1 Introduction 102
7.2 Analysis 103
7.3 Conclusion 105
References 105
Chapter 8: Implementation and Assessment of High-Throughput Donor Typing at the Milan, Italy, Immunohematology Reference Laboratory 106
8.1 Introduction 106
8.2 BeadChip™ System Implementation 107
8.3 Results 108
8.3.1 HPA BeadChipTM 108
8.3.2 HEA BeadChipTM 108
8.4 Results Following Routine Implementation of HEA and HPA BeadChip™ Analysis 109
8.5 Conclusion 110
References 111
Chapter 9: Implementation of the BioArray Human Erythrocyte Antigen (HEA) BeadChip™ System: The Spanish Red Cross Blood Centre of Madrid, Spain Experience 112
9.1 Introduction 112
9.1.1 History and service of the Red Cross Blood Centre of Madrid 112
9.2 Blood Group Classification 113
9.3 The BioArray BeadChip™ System 114
9.3.1 HEA BeadChipTM Assay 114
9.3.2 HEA BeadChip Training 115
9.3.3 HEA BeadChip Materials 115
9.3.4 Implementation of HEA BeadChip: DNA Extraction 116
9.4 Validation of DNA Extraction Technique 116
9.5 Initial Results of HEA BeadChip™ Array Implementation 117
9.6 Future Objectives 117
References 118
Chapter 10: Looking Beyond HEA: Matching SCD Patients for RH Variants 120
10.1 Background 121
10.2 Transfusion Therapy for Patients with Sickle Cell Disease 122
10.2.1 The Stroke Prevention Trials 122
10.2.2 Transfusion Therapy in Alloimmunized Patients with SCD 122
10.3 DNA Testing for the Prediction of Blood Groups 124
10.3.1 Genes Encoding Blood Groups 125
10.3.2 Experience with High-Throughput DNA Arrays 125
10.4 DNA Testing for Prediction of RH Alleles 126
10.5 Testing with RHD BeadChip™ and RHCE BeadChip™ 129
10.5.1 Test Samples 131
10.5.2 Analyses 131
10.5.3 Interpretation of Set #1: Interpretation of Samples from African-Americans with Known Variant RH Alleles (Diverse Samples) 131
10.5.3.1 RHD Analysis in Set#1 131
10.5.3.2 RHCE Analysis in Set#1 132
10.5.4 Interpretation of Set #2: Interpretation of Samples from Random African-American Donors and Patients with SCD 132
10.5.4.1 RHD Analysis in Set #2 132
10.5.4.2 RHCE Analysis in Set #2 133
10.6 Conclusions and Perspectives 134
References 137
Chapter 11: Identification of Altered RHD and RHCE Alleles: A Comparison of Manual and Automated Molecular Methods 140
11.1 Background 141
11.1.1 RHD 141
11.1.2 RHCE 142
11.1.3 Automation 142
11.2 Methods 143
11.2.1 Manual Assays 143
11.2.2 Automated RHD and RHCE BeadChip™ 144
11.3 Results 144
11.3.1 RHD BeadChip™ 144
11.3.2 RHCE BeadChip 145
11.4 Conclusion 148
References 150
Chapter 12: Bayesian Classification Algorithms for Automated Allele Assignment 151
12.1 Introduction 151
12.2 Bayesian Classification for Automated Allele Assignment 153
12.3 Bayesian Analysis of the HLA Complex 157
12.3.1 Methodology 158
12.3.2 Classifier Training by Supervised Learning 158
12.3.3 Thresholds and Transition Probabilities 159
12.3.4 Allele-Pair Assignment 160
12.4 Bayesian RHCE and RHD Variant Analysis 160
12.4.1 Methodology 160
12.4.2 Error Rates 162
12.4.3 RHCE Variant Analysis 163
12.4.4 RHD Variant Analysis: Neural Network Classification of Deletions and Hybrids 163
12.5 Implementation 165
12.6 Summary 165
References 165
Index 167