Practical Flow Cytometry in Haematology Diagnosis (eBook)

Allyson Doig, Mark Drummond, Mike Leach (Autoren)

eBook Download: PDF

2013
Wiley (Verlag)
9781118487990 (ISBN)

Infos

The accurate diagnosis of haematologic malignancies is a complex and challenging task. It routinely involves morphologic, molecular, cytogenetic and flow cytometric expertise. To determine what treatment protocol will be followed, it is vital to integrate, interpret and report these results accurately.

Flow cytometry is key in this diagnostic pathway.

This book guides the reader as to how flow cytometry results should be interpreted and applied to optimize patient care. At the core of this text is an appreciation of clinical, morphological and immunophenotypic correlation and the importance of constant liaison and discussion between the medical and scientific teams.

The authors present a logical and practical approach to the diagnosis of blood disorders (both neoplastic and reactive) and evaluate the diagnostic applications of flow cytometry.

Practical Flow Cytometry in Haematology Diagnosis provides:

• A clinical reference source on all aspects of flow cytometry, covering both malignant and benign conditionsCarefully chosen real-life cases in each chapter, complemented by high quality morphological images
• Help in making a diagnosis, together with an understanding of the limitations of the technique and the potential pitfalls

All those who instigate, perform, interpret or act upon flow cytometry patient material will find this book an invaluable guide.

Dr Mike Leach - Department of Haematology, Gartnavel General Hospital, Glasgow, UK

Dr Mark Drummond - Haematology Department, Beatson West of Scotland Cancer Centre, Glasgow, UK

Dr Allyson Doig - Department of Haematology, Gartnavel Hospital, Glasgow, UK

Inhalt

Practical Flow Cytometry in Haematology Diagnosis 5
Copyright 6
Contents 7
Preface 9
Acknowledgements 10
Chapter 1 Introduction 11
References 12
Chapter 2 Principles of Flow Cytometry 13
Introduction 13
Sample preparation 13
Direct staining method for surface immunophenotyping of peripheral blood or bone marrow (lyse – no wash method) 13
Surface immunoglobulin staining 14
Intracellular Staining 14
The flow cytometer 14
The fluidic system 14
The optical system 15
Light scatter 15
Fluorescence 16
Fluorescence intensity 18
Spectral overlap 18
The electronic system 19
Threshold 19
Data display 19
Histogram 21
Dot plot 21
Instrument set-up and quality control 25
Acknowledgements 28
References 29
Chapter 3 Limitations 30
Introduction 30
Clinical context issues 30
Sampling issues 30
Blood 30
Bone marrow aspirates 31
Effusions 33
Cerebrospinal fluid 33
Regenerating bone marrows 35
Technical issues 35
Operator training 35
Antibody and fluorochrome selection 37
Background fluorescence 37
Interpretation issues 38
Morphological correlation 38
Clinical context 38
Combined report 38
Conclusion 39
References 39
Chapter 4 Normal Blood and Bone Marrow Populations 41
Normal stem and precursor cell populations 41
Haematopoietic progenitor cells 41
Myeloid maturation 43
Erythroid maturation 45
Megakaryocytic maturation 45
Lymphoid maturation 45
Haematogones 46
References 51
Chapter 5 Acute Leukaemia 53
Introduction 53
Acute leukaemia: definition and classification 53
Clinical presentation and sampling for flow cytometry 54
Identification of neoplastic precursor cells 55
The blast population 55
Blast populations are heterogeneous 55
Correlating the immunophenotype with genetic analysis 56
Blast cell characteristics 56
Patterns of precursor antigen expression in acute leukaemia 60
Enumeration of blast cells 61
Lineage assignment of acute leukaemia 62
Lineage assignment in acute myeloid leukaemia 62
Lineage assignment in acute lymphoblastic leukaemia 63
Acute leukaemia of ambiguous lineage 63
Acute myeloid leukaemia 64
Aberrant and asynchronous antigen expression in acute myeloid leukaemia 65
Antigenic patterns of particular significance in acute myeloid leukaemia 76
CD34 -negative HLA-DR-negative acute myeloid leukaemia 76
Identifying particular acute myeloid leukaemia subtypes by flow 77
Precursor lymphoid neoplasms (acute lymphoblastic leukaemia/lymphoma) 87
B-cell acute lymphoblastic leukaemia/lymphoma 91
Aberrant antigen expression in B-acute lymphoblastic leukaemia 93
Identifying B-acute lymphoblastic leukaemia subtypes by flow 93
T-cell acute lymphoblastic leukaemia/lymphoma 94
Aberrant antigen expression in T-acute lymphoblastic leukaemia 96
Examination of cerebro-spinal fluid and serous effusions 98
Central Nervous System Disease in Acute Leukaemia 98
Detection of acute leukaemia cells in serous effusions 99
Identification of genetic abnormalities by flow 102
Conclusion 103
References 103
Chapter 6 Chronic Lymphoid Leukaemias and Exfoliating Lymphoma 110
Normal peripheral blood lymphoid populations 110
Identification of clonal lymphoid populations 111
Identification of clonal B-cell disorders 111
Diagnosis of B-cell disorders 111
CD5 positive B-cell lymphoproliferative disorders 112
Chronic lymphocytic leukaemia 112
Monoclonal B lymphocytosis 115
Mantle cell lymphoma 115
Splenic marginal zone lymphoma 117
B Prolymphocytic leukaemia 118
Lymphoplasmacytoid lymphoma 120
Diffuse large B-cell lymphoma, Richter type 123
CD10 Positive B-cell lymphoproliferative disorders 126
Follicular lymphoma 126
Diffuse large B-cell lymphoma 128
Burkitt lymphoma 128
Precursor B acute lymphoblastic leukaemia 129
Hairy cell leukaemia 131
B-cell disorders identified using an extended B-cell panel 131
Hairy Cell Leukaemia 132
Hairy Cell leukaemia variant 133
Splenic Marginal Zone Lymphoma 133
Identification of clonal T-cell disorders 135
Diagnosis of specific T-cell disorders 137
CD4 Positive T-cell disorders 138
Sezary syndrome/cutaneous T-cell lymphoma 138
T-cell prolymphocytic leukaemia 139
Acute T-cell leukaemia/lymphoma 141
Anaplastic large cell lymphoma 142
Angio-immunoblastic T-cell Lymphoma 143
CD8 Positive T-cell disorders 146
T-cell large granular lymphocyte leukaemia 146
Natural killer cell chronic lymphoproliferations 147
Hepatosplenic T-cell lymphomas 149
T-prolymphocytic leukaemia 151
CD4/CD8 positive T-cell disorders 151
CD4/8 Negative disorders 151
Aggressive natural killer/T-cell neoplasms 151
Enteropathy associated T-cell lymphoma 152
Hepatosplenic T-cell lymphomas 155
Peripheral T-cell lymphoma, not otherwise specified 155
CD30 positive tumours 155
Flow cytometric assessment of serous effusions 156
Primary effusion lymphoma 156
Flow cytometric assessment of cerebrospinal fluid 156
Identification of overt central nervous system disease 157
Identification of subclinical disease 157
Summary 159
References 159
Chapter 7 Myelodysplastic Syndromes and Myeloproliferative Neoplasms 162
Introduction 162
Myelodysplastic syndromes 162
Classification of myelodysplastic syndromes 165
Prognosis of myelodysplastic syndromes 165
Flow cytometry in myelodysplastic syndromes 165
Myelodysplastic/myeloproliferative neoplasms 173
Chronic myelomonocytic leukaemia 173
Chronic myeloid leukaemia 175
The BCR-ABL negative myeloproliferative neoplasms 176
Mastocytosis 177
Summary 180
References 180
Chapter 8 Disorders of Plasma Cells 183
Plasma cell disorders 183
Current diagnostic criteria 183
Monoclonal gammopathy of undetermined significance 183
Smouldering myeloma 184
Multiple myeloma 184
Plasma cell leukaemia 184
Plasmacytoma 184
Primary amyloidosis 184
Plasma cell morphology 185
The applications of immunophenotyping to plasma cell disorders 188
Prognosis 190
References 193
Chapter 9 Minimal Residual Disease 194
Introduction 194
Chronic lymphocytic leukaemia 195
Acute myeloid leukaemia 196
Minimal residual disease analysis in the management of acute lymphoblastic leukaemia 197
Monitoring minimal residual disease in acute lymphoblastic leukaemia 197
Minimal residual disease flow immunophenotyping versus molecular polymerase chain reaction analysis 199
Technical aspects of minimal residual disease flow assessment in acute lymphoblastic leukaemia 199
Technical challenges 203
Conclusions 209
Acknowledgements 210
References 210
Chapter 10 Red Cells, Leucocytes and Platelets 212
Paroxysmal nocturnal haemoglobinuria 212
Indications for paroxysmal nocturnal haemoglobinuria screening 214
Analysis of red cells 214
Analysis of granulocytes 215
Analysis of monocytes 215
Analysis of lymphocytes 216
Analysis of platelets 216
Paroxysmal nocturnal haemoglobinuria clones in myelodysplastic syndromes 217
Summary 218
Red cell membrane disorders 218
Foetal maternal haemorrhage 221
Lymphocyte subset analysis and immunodeficiency 224
Haemopoietic stem cell enumeration 224
Granulocyte disorders 225
Platelet disorders 229
Inherited disorders 229
Acquired disorders 230
Monitoring of antiplatelet drug therapy 230
References 231
Chapter 11 Reactive and Non-neoplastic Phenomena 235
Peripheral blood 235
Lymphocytosis 235
Lymphopenia 239
Bone marrow 241
Pleural and pericardial fluids 246
Bronchoalveolar lavage specimens 248
Cerebrospinal fluid 248
References 250
Index 253

Rezensionen

"I would particularly recommend this book to haematology
trainees, as an introduction to, but not as a replacement for, a
period spent in an immunophenotyping laboratory."
(British Journal of Haematology, 14 November 2014)

"There is no doubt that this book will find its way firmly
onto the bookshelf in every laboratory, and into the work bags of
many trainees." (Haem Trainee, 1 January
2014)

"This is a concise and very practical guide to the use of
flow cytometry in hematological and hematopathological diagnosis.
While not as detailed in immunophenotyping intricacies as other
books of its kind, it shines with its practical approach and expert
guidance applied to current clinical practice."
(Doody's, 19 July 2013)

Produkt-Details

• Erscheint lt. Verlag: 30.1.2013
• Sprache: englisch
• Themenwelt: Medizinische Fachgebiete ► Innere Medizin ► Hämatologie
• Themenwelt: Studium ► 2. Studienabschnitt (Klinik) ► Anamnese / Körperliche Untersuchung
• Schlagworte: Accurate • Book • Care • Clinical • Complex • Core • cytometric • Cytometry • determine • Diagnosis • Durchflusszytometrie • Expertise • flow • Hämatologie • Hämatologisches Labor • Haematologic • Hämatologie • Hämatologisches Labor • Interpret • Key • laboratory hematology • Malignancies • Medical Science • Medizin • Morphologic • optimize • pathway • Patient • Reader • Text • treatment protocol • Vital
• ISBN-13: 9781118487990 / 9781118487990