Dacie and Lewis Practical Haematology: Expert Consult: Online and Print

Front matter
......Page 1
Copyright
......Page 4
Preface
......Page 5
Contributors
......Page 7
Outline placeholder......Page 0
Specimen Containers......Page 11
Waste Disposal......Page 12
Blood Film Preparation......Page 13
Defibrinating Whole Blood......Page 14
Ethylenediaminetetra-acetic Acid (EDTA)......Page 15
Effects of storage on blood cell morphology......Page 16
References......Page 18
Reference ranges......Page 19
Confidence Limits......Page 20
Normal reference values......Page 21
Quality assurance procedures......Page 588
Availability of tests at each level......Page 81
Combined Warm and Cold Autoantibodies......Page 251
`Essential´ haematology tests......Page 26
Acetic acid......Page 111
Neutral Buffered EDTA, pH 7.0......Page 524
Antigens......Page 27
Precision......Page 28
References......Page 599
Basic haematological techniques......Page 31
Measurement of haemoglobin concentration using a spectrometer (spectrophotometer) or photoelectric colorimeter......Page 32
Quantitative Abnormalities of Blood Cells......Page 33
Preparation of Standard Graph and Standard Table......Page 34
Reagents......Page 35
Care of the Microscope......Page 283
Tissue Factor......Page 400
Method......Page 36
Calibration Audits......Page 311
Youden (xy) plot......Page 595
Rapid Staining Method......Page 37
Waterbaths......Page 404
Manual differential leucocyte count......Page 38
Method......Page 39
Reference Differential White Cell Count......Page 40
Method......Page 41
Differentiating between Reticulocytes and Other Red Cell Inclusions......Page 43
Automated blood count techniques......Page 44
Impedance Counting......Page 45
Reliability of electronic counters......Page 46
Packed cell volume and mean cell volume......Page 47
Variations in red cell volumes: red cell distribution width......Page 49
Total white blood cell count......Page 50
Automated differential count......Page 51
Automated digital imaging analysis of blood cells......Page 52
Automated instrument graphics......Page 53
Mean Platelet Volume......Page 55
Immature Reticulocyte Fraction......Page 56
Calibration of automated blood cell counters......Page 57
Flagging of automated blood counts......Page 58
Microscope Components......Page 59
Routine Maintenance of the Microscope......Page 60
References......Page 61
Manual Method......Page 65
Fixing Blood Films......Page 66
Lymphocytosis......Page 67
Diagnostic Reliability......Page 604
Azure B-Eosin Y Stock Solution......Page 68
Determination of the International Normalized Ratio......Page 69
Antibodies......Page 402
External Quality Assessment......Page 188
Allele-Specific Oligonucleotide Hybridization......Page 70
Negative control......Page 223
Staining of Sections of Bone Marrow Trephine Biopsy Specimens......Page 71
Interpretation......Page 346
Utility of the Buffy Coat......Page 72
Sizes of tubes......Page 620
Principle......Page 455
Parasites detectable in blood, bone marrow or splenic aspirates......Page 73
Results and Interpretation......Page 349
Classification of Acute Myeloid Leukaemia......Page 74
Interlaboratory Communication......Page 261
References......Page 75
Blood cell morphology in health and disease......Page 77
Preparation of commonly used reagents......Page 78
Abnormal erythropoiesis......Page 79
Standardization of Oral Anticoagulant Treatment......Page 237
Hypochromia (Hypochromasia) (πρ, under)......Page 82
Hyperchromasia (Hyperchromia) (περ, over)......Page 84
β Thalassaemia Syndromes......Page 310
Irregularly Contracted Red Cells......Page 85
Method......Page 343
Schistocytosis (Fragmentation) (σχiotaστzeta, cleft)......Page 88
Therapeutic Range and Choice of Thromboplastin......Page 238
DAT-Negative Autoimmune Haemolytic Anaemia......Page 286
Principles of Laboratory Analysis......Page 90
Acquired B......Page 91
Internal audit for total quality management......Page 453
Abnormalities of All Cell Lines......Page 552
Pappenheimer Bodies......Page 93
Post-Analytical Phase......Page 570
Interpretation......Page 94
Erythroblastaemia......Page 95
Principle......Page 96
Method15......Page 97
Thrombophilia......Page 154
Preparation of QC Material for Platelet Count......Page 100
Pelger-Huët Cells......Page 101
Eosinophils......Page 102
Universal Precautions......Page 579
Lymphocytes......Page 103
Tube Tests......Page 105
References......Page 178
Supplementary techniques including blood parasite diagnosis......Page 109
Standardization......Page 585
Method......Page 110
Reference Standards......Page 210
Warm Autoantibodies......Page 281
Level C......Page 603
Mechanism of Erythrocyte Sedimentation......Page 112
General approach to investigation of haemostasis......Page 113
Haemolysates......Page 239
Demonstration of Lupus Erythematosus Cells......Page 114
Method......Page 115
Method4,15......Page 189
Phosphate Buffer, Sörensen's......Page 619
Kidd antibodies......Page 151
Principles of parasite detection......Page 116
Range of white blood cell count in health......Page 475
Staining Thin Films......Page 117
Method......Page 477
Rapid diagnostic tests for malaria......Page 118
Method......Page 124
Capillary tube method......Page 125
Method......Page 194
Hirudin......Page 391
Determination of Sites of Red Cell Destruction Using 51Cr......Page 353
Antenatal serology and haemolytic disease of the newborn......Page 126
References......Page 127
Bone marrow biopsy......Page 130
Diagnostic radioisotopes in haematology......Page 377
Haemoglobins with Reduced Solubility......Page 131
ABO and D grouping......Page 132
Platelets......Page 398
Blood Coagulation......Page 133
Eosinophilia......Page 149
Calculation of International Sensitivity Index......Page 470
Principles of Marrow Aspirate Examination......Page 135
Interpretation......Page 451
Rh Antigens and Encoding Genes......Page 489
Sources of Error and Physiological Variations......Page 136
Higher Power (x40, x100 Oil-Immersion)......Page 137
Data processing......Page 568
HEPES Buffer, pH 6.6......Page 618
Percutaneous trephine biopsy of the bone marrow......Page 138
Processing of Bone Marrow Trephine Biopsy Specimens......Page 140
References......Page 144
Introduction to the analysis of DNA......Page 145
Principle......Page 146
Citrate-Phosphate-Dextrose-Adenine (CPD-A) Solution, pH 5.6-5.8......Page 147
Method......Page 150
Method......Page 152
Aplastic Anaemia11......Page 386
Laboratory Safety......Page 155
Cytogenetic Principles and Terminology......Page 156
Fluorescence in situ Hybridization......Page 159
Translocations, Molecular Analysis and Minimal Residual Disease......Page 160
Interpretation of Survival Curves......Page 161
Electronic Issue......Page 162
cDNA synthesis......Page 163
Indirect Donath-Landsteiner Test......Page 164
Advantages of qRT-PCR......Page 165
Reagents......Page 167
Method......Page 324
Antiplatelet therapy......Page 169
Interpretation......Page 170
Addition of red cell suspensions to dilutions of serum......Page 503
Acknowledgements......Page 230
Principle: JAK2 Exon 14 Mutation Analysis......Page 171
CaCl2......Page 173
11q23 Abnormalities......Page 174
Paroxysmal nocturnal haemoglobinuria......Page 296
Principle......Page 175
Reagents and method......Page 461
Flow Cytometry Analysis of the GPI-Linked Proteins on Red Cells......Page 176
Interpretation......Page 177
Iron deficiency anaemia and iron overload......Page 181
Technology and automation in blood transfusion laboratories......Page 519
Dietary Iron Absorption......Page 182
Method......Page 469
Types of Autoantibody......Page 280
Red Blood Cell Lysing Methods......Page 183
Anti-A1 and anti-H......Page 184
Methods for assessing iron status......Page 185
Principle......Page 284
Quality Control of a Test Method (Technical Quality)......Page 186
Point-of-Care Testing......Page 472
Direct Reading Haemoglobinometers......Page 606
Method......Page 288
Calculation......Page 191
Microcytic Hypochromic Anaemias......Page 224
Quality Assurance within the Laboratory......Page 225
Significance of Methaemalbuminaemia......Page 192
Tubes......Page 456
Indirect Antiglobulin Crossmatch......Page 531
Method......Page 193
Macroscopic reading......Page 498
Specimen Shipping......Page 245
Transferrin Index......Page 195
Assessment of Fetal Anaemia......Page 460
Assays for the Serum Transferrin Receptor......Page 196
Isoantibodies......Page 197
Analysers......Page 198
Autoantibodies......Page 412
Methodological and biological variability of assays......Page 199
Thromboplastin......Page 200
Functional Iron Deficiency......Page 201
Chronic Transfusion Programmes......Page 202
Investigation of megaloblastic anaemia......Page 207
Folate absorption and metabolism......Page 208
Differential Diagnosis of Macrocytic Anaemia......Page 211
Unstained preparations......Page 341
Sensitivity and Specificity of Cobalamin and Holotranscobalamin Assays......Page 216
Low Ionic Strength Solution (LISS)1......Page 252
Counting Technique......Page 217
Clinical and Diagnostic Pitfalls of Folate Assays......Page 219
Principle......Page 381
Genetic Factors......Page 220
Maintaining quality and reliability of tests......Page 221
Management of Overanticoagulation......Page 222
Normal Range of Autohaemolysis......Page 226
ANOVA......Page 622
Classification of haematological neoplasms......Page 555
Calculation......Page 227
Compatibility Testing......Page 228
References......Page 613
Antenatal Serology......Page 229
References......Page 231
Laboratory methods used in the investigation of the haemolytic anaemias......Page 235
Investigation of haemolytic anaemia......Page 236
Monocytosis......Page 548
Haemoglobinometry......Page 605
Linearity......Page 567
Laboratory Control of Heparin Treatment......Page 473
Preparation of glassware......Page 405
Normal Ranges......Page 240
Examination of plasma (or serum) for methaemalbumin......Page 241
Method......Page 320
Preparation of liquid suspension......Page 242
Control Plasma......Page 243
Method......Page 244
Waste Disposal......Page 581
Gloves......Page 479
Spectroscopic Examination of Blood for Methaemoglobin and Sulphaemoglobin......Page 246
Method......Page 247
Testing Fetal DNA in the Maternal Circulation......Page 263
Method......Page 248
References......Page 249
Investigation of the hereditary haemolytic anaemias......Page 250
Level B: District Hospitals......Page 522
Control Charts......Page 590
Buffers......Page 255
Imidazole Buffered Saline, pH 7.4......Page 256
Method......Page 608
Defibrinating Blood......Page 257
Platelet Count......Page 317
Preparation of Stable Control Material for EQAS......Page 318
Preparation of Surrogate Leucocytes......Page 598
Reagents for Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT)......Page 258
Detection of enzyme deficiencies in hereditary haemolytic anaemias......Page 259
Use of Enzyme-Treated Cells......Page 260
Sodium hypochlorite (chlorine)......Page 580
Monospecific reagents......Page 262
Alloantibodies......Page 504
Interpretation......Page 264
Reagent......Page 299
Measurement of Hb A2 by Microcolumn Chromatography with Tris-HCl Buffers......Page 265
Calculation of Enzyme Activity......Page 266
Interpretation of Results......Page 267
Method......Page 268
Principle......Page 269
Normal Range......Page 270
Measurement of Red Cell 2,3-Diphosphoglycerate......Page 271
Reaction......Page 272
Oxygen dissociation curve......Page 273
Interpretation......Page 274
References......Page 275
Assessment of the blood film and count in suspected acquired haemolytic anaemia......Page 278
Erythrocytes......Page 468
Immune haemolytic anaemias......Page 279
Methods of Investigation......Page 282
Geometric Mean Normal Prothrombin Time......Page 285
Capillary Reagent......Page 471
Continuous Quality Improvement......Page 287
Red Cell Disorders......Page 553
Elution of Antibodies from Red Cells......Page 289
Myelomatosis (Plasma Cell Myeloma)......Page 291
Oral anti-IIa and anti-Xa agents......Page 292
Specificity of the Donath-Landsteiner antibody......Page 293
Drug-Induced Autoimmune Haemolytic Anaemias......Page 294
Note......Page 295
Blood Count and Film......Page 326
Alloantibodies......Page 507
Principle......Page 298
Method......Page 300
Acute Transfusion Reactions......Page 301
Specimen Collection......Page 462
References......Page 302
Investigation of abnormal haemoglobins and thalassaemia......Page 306
Structural variants of haemoglobin......Page 307
Internal Quality Control......Page 308
Thalassaemia syndromes......Page 309
Measurement of radioactivity......Page 362
Reductions in more than one cell line......Page 312
Accuracy and Comparability......Page 313
Preparation of Haemolysate for Qualitative Haemoglobin Electrophoresis......Page 314
Cellulose Acetate Electrophoresis at Alkaline pH......Page 315
Mean Normal Plasma Volume (ml)......Page 384
Interpretation and Comments......Page 316
Protamine Neutralization Test......Page 371
Modified (One-Tube) Osmotic Fragility Test......Page 319
Phospholipid Reagent for Activated  Partial Thromboplastin Time......Page 611
Reagents......Page 322
Detection of an unstable haemoglobin......Page 323
Massive Transfusion......Page 533
Differential diagnosis of common haemoglobin variants......Page 325
Method......Page 327
Reagents......Page 328
Method......Page 329
Investigation of a transfusion reaction......Page 541
Quantitation of Hb F......Page 330
Interpretation and Comments......Page 331
Assessment of the intracellular distribution of Hb F......Page 332
Assessment of iron status in thalassaemia......Page 333
Interpretation and Comments......Page 334
References......Page 335
Erythrocyte cytochemistry......Page 338
Initial screening tests......Page 547
Method of Staining Siderotic Granules......Page 339
Detection of membrane antigens......Page 359
Standardization of Working Conditions......Page 342
Standardization of EQA Schemes......Page 593
Method......Page 345
Methodology check......Page 382
Results and Interpretation......Page 347
Haemolytic Anaemias......Page 348
Naphthol AS-D Chloroacetate Esterase......Page 350
Technical considerations......Page 351
Alcohols......Page 352
Antibody Quantitation......Page 354
References......Page 355
Introduction......Page 358
Acid-Citrate-Dextrose (ACD) Solution - NIH-A......Page 397
Method 1: Wash-Stain-Lyse-Wash......Page 361
Quantification of Antigens......Page 363
Barbitone Buffer, pH 7.4......Page 525
Quantification: calculation of ABC or MESF values......Page 364
Panel of McAb Useful for Diagnosis and Classification......Page 366
Principle......Page 385
Panel of McAb for Diagnosis and Classification......Page 369
CD38 and ZAP70 expression......Page 370
References......Page 375
Radiation protection......Page 378
Level C: Central and Teaching Hospitals......Page 380
Qualitative Abnormalities of Blood Cells......Page 383
Radioactive Chromium (51Cr) Method......Page 387
Calculations......Page 388
Correction for Elution......Page 389
Miscellaneous Imaging......Page 392
Fetal Blood Sampling......Page 393
References......Page 394
Investigation of haemostasis......Page 396
Secretors and Non-Secretors......Page 487
Methods......Page 523
Platelet Aggregation......Page 399
Cofactors......Page 401
Clinical Approach......Page 403
Storage of Sera or Plasma......Page 406
Storage of Plasma and Sample Thawing......Page 407
Laboratory support for management of hiv/aids: cd4-positive t-cell counts......Page 408
Percentage Detection Method......Page 409
Analysis Time Over......Page 410
Molecular Mechanism of the Biphasic Waveform: LC-CRP......Page 411
Principle......Page 413
Delayed Haemolytic Transfusion Reaction......Page 414
Interpretation......Page 415
3......Page 416
7......Page 418
Reagents......Page 419
General Principles of Parallel Line Bioassays of Coagulation Factors......Page 420
Reagents......Page 421
Calculation of Results......Page 422
Monitoring Replacement Therapy in Coagulation Factor Defects and Deficiencies......Page 423
Method......Page 424
Method......Page 425
Tests for Other Inhibitors......Page 426
Bleeding Time......Page 427
Reagents......Page 428
von Willebrand Factor Antigen Immunoturbidimetric Assay......Page 429
Standard Curve......Page 430
Assay Method......Page 431
Running buffer......Page 432
Electrophoresis (Day 1 Evening)......Page 433
Laboratory Investigation of Platelets and Platelet Function......Page 434
Principle......Page 435
Aggregating agents......Page 436
Arachidonic acid......Page 437
Adenosine 5'-diphosphate......Page 438
Normal Range......Page 439
Further Investigation of Platelet Function......Page 441
Principle......Page 442
Method......Page 443
Family Studies......Page 444
Genotype Assignment......Page 445
Introduction to thrombophilia......Page 449
Sample Preparation......Page 450
Interpretation of Tests for Lupus Anticoagulant......Page 452
Antithrombin (AT)......Page 454
Principle......Page 457
Staff Training......Page 612
Controls......Page 458
Investigation of a Patient Who Bleeds While Taking Thrombolytic Agents or Immediately Afterwards......Page 480
Investigation of the Fibrinolytic System: General Considerations......Page 459
Homocysteine......Page 463
Global assays of coagulation......Page 464
ABO Antigens and Encoding Genes......Page 486
Therapeutic Range......Page 474
Heparin-Induced Thrombocytopenia......Page 476
Diamed Heparin-PF4 Antibody Test......Page 478
Red Cell Antigens......Page 483
ABO System......Page 485
Lewis Antigens and Encoding Genes......Page 488
Antibodies......Page 490
Packed Cell Volume......Page 491
Macrocytic Anaemias......Page 492
Antigen-Antibody Reactions......Page 495
Accreditation......Page 575
Neutropenia......Page 496
Reagent Red Cells......Page 497
Demonstration of Lysis......Page 499
Autodiluters......Page 629
Quality Control of Antiglobulin Reagents......Page 500
Recommended Antiglobulin Test Procedure......Page 501
Assessment of Individual Worker Performance......Page 502
Autoantibodies......Page 506
Methods of Demonstrating Antibodies......Page 508
Granulocyte Preparation......Page 509
Selection of Blood and Other Components......Page 539
Indirect Test......Page 510
Chloroquine Treatment of Platelets and Granulocytes......Page 511
MAIPA Assay......Page 512
Molecular Genotyping of Platelet Alloantigens......Page 513
References......Page 514
Laboratory aspects of blood transfusion......Page 518
Pre-transfusion compatibility systems......Page 521
Point-of-Care Testing......Page 526
Methods......Page 527
Controls......Page 528
Standardized Counting Chambers......Page 609
Method......Page 529
Selection and transfusion of red cells......Page 530
False-positive results (in immediate spin crossmatch)......Page 532
Follow-Up Antibody Screening......Page 534
Antenatal Assessment of the Severity of Haemolytic Disease of the Fetus and Newborn......Page 535
Tests on Maternal and Cord Blood at Delivery......Page 536
Anti-D Prophylaxis......Page 537
Recommended Action at Delivery (or Potentially Sensitizing Event)......Page 538
Patients Receiving Transfusions at Close Intervals......Page 540
Acute Intravascular Haemolysis......Page 542
Serological Investigation......Page 543
Microcytic Anaemia......Page 549
Deviation index......Page 594
Macrocytic Anaemia......Page 551
Benchmarking......Page 554
Classification of the Myelodysplastic Syndromes......Page 556
Classification of Myeloproliferative Neoplasms and Related Conditions......Page 557
Laboratory organization and management......Page 561
Staff Appraisal......Page 562
Water......Page 563
Calculation of Test Costs......Page 564
Test selection......Page 565
Test Utility......Page 566
Laboratory Computers......Page 569
Method......Page 572
Method......Page 573
Audit......Page 574
Principles of Safety Policy......Page 577
Electrical and Radiation Safety......Page 578
References......Page 583
Control materials and reference standards......Page 586
Correlation Check......Page 591
External Quality Assessment......Page 592
Interpretive Tests......Page 596
Preparation of Preserved Whole Blood......Page 597
Introduction: types of laboratories......Page 601
Level A: Sub-District Facilities Including Health Centres......Page 602
Counting Chambers......Page 607
Peripheral Blood Morphology......Page 610
Further Useful Publications......Page 614
Sources of Teaching Material and Equipment for Under-resourced Laboratories......Page 615
Acid-Citrate-Dextrose (Alsever's) Solution......Page 616
Neutral EDTA, pH 7.0, 110mmol/l......Page 617
Statistical procedures......Page 621
Automated (mechanical) pipettes......Page 624
A
......Page 630
B
......Page 632
C
......Page 634
D
......Page 635
E
......Page 636
F
......Page 637
H
......Page 638
I
......Page 640
L
......Page 641
M
......Page 642
N
......Page 643
P
......Page 644
R
......Page 646
S
......Page 647
T
......Page 648
Z
......Page 650