The science of laboratory diagnosis

John Crocker, David Burnett0470859121, 9780470859124

This fully revised and updated edition of The Science of Laboratory Diagnosis provides a concise description of all common laboratory tests available in medical practice with notes on their application, the accuracy of each test, the historical background to the adoption of various tests and their effectiveness in diagnosis. Well illustrated, with clear headings, tables, flow charts and pathology slides, most in full colour Provides an accessible reference book in which relevant information can be found easily Page design facilitates rapid assimilation of principles and key facts All the chapters have been updated and new material has been introduced to cover recently developed techniques, such as fluid-based cytology, telepathology and proteomics
The Science of Laboratory Diagnosis, Second Edition is an essential primary reference source for everyone working in a clinical laboratory. This book is essential reading for pathologists, biomedical scientists, medical laboratory scientific officers and all clinicians involved in laboratory research.
Reviews of the First Edition:
” The text is concise, wide-ranging and easy to digest. The ease of extraction of the important facts make it an ideal source of information for use in a variety of situations from the postgraduate examination to the clinical directors’ board meeting.” BULLETIN OF THE ROYAL COLLEGE OF PATHOLOGISTS
” The editors have done a marvellous job, more than fulfilling their stated aim of producing a volume describing the multidisciplinary state of modern pathology which will be of interest to a wide range of readers. … I was particularly impressed by the many tables and flow charts, which can be used as aids to decision making .” JOURNAL OF CLINICAL PATHOLOGY
” This is an excellent book to dip into and get a feel for techniques used in the other disciplines of pathology. ” ANNALS OF CLINICAL BIOCHEMISTRY

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Table of contents :
THE SCIENCE OF LABORATORY DIAGNOSIS Second Edition……Page 3
Contents……Page 9
Preface to the first edition……Page 17
Preface to the second edition……Page 19
Contributors list……Page 21
SECTION 1 HISTOPATHOLOGY……Page 25
Introduction……Page 27
Specimen receipt and handling……Page 28
Fixation and decalcification……Page 29
Tissue processing and embedding……Page 31
Microtomy……Page 35
Special techniques……Page 37
Acknowledgements……Page 39
Basic staining mechanisms……Page 41
Common staining methods……Page 44
Acknowledgements……Page 49
Structure and classification……Page 51
Preservation and preparation……Page 52
Histochemical techniques……Page 53
Diagnostic applications of enzyme histochemistry……Page 55
References……Page 58
Multi-stage methods……Page 59
Imaging……Page 65
Further reading……Page 66
Introduction……Page 67
Principles of EM technique and organization……Page 68
Applications of transmission electron microscopy……Page 72
Specialized techniques and procedures……Page 78
Acknowledgements……Page 83
Further reading……Page 84
Basic microscope design……Page 85
Types of microscopy……Page 87
Photomicrography……Page 90
Further reading……Page 91
Methods……Page 93
Applications of measurement in diagnostic pathology……Page 99
Automation in tissue histology……Page 103
Further reading……Page 105
Proliferation markers……Page 107
Practical use of markers……Page 112
Method of evaluation……Page 113
Further reading……Page 116
SECTION 2 CYTOLOGY……Page 117
Types of cytology specimen……Page 119
Cell concentration techniques……Page 123
Staining methods……Page 127
Cytodiagnosis……Page 128
Cytomorphology……Page 129
Accuracy and limitations of cytology……Page 136
Gynaecological cytopathology and cervical screening……Page 137
Further reading……Page 143
SECTION 3 MICROBIOLOGY – BACTERIOLOGY……Page 145
Fluorescence microscopy……Page 147
Examination of unstained specimens by transmitted light……Page 148
Staining of specimens……Page 149
Further reading……Page 152
Culture media in microbiology……Page 153
Formulation of culture media……Page 154
Chemically defined culture media versus complex undefined media……Page 155
Storage of culture media……Page 156
Quality tests……Page 157
Further reading……Page 158
Direct identification……Page 159
Identification following culture……Page 160
Identification of bacteria of medical importance……Page 164
Further reading……Page 168
Blood culture systems……Page 169
Antimicrobial susceptibility testing and identification systems……Page 170
Urine screening systems……Page 172
Identification and antimicrobial susceptibility testing systems for mycobacteria……Page 173
Further reading……Page 174
Sample preparation……Page 175
Amplification……Page 176
Current practice……Page 177
Future application……Page 180
Further reading……Page 181
Tests for antibody……Page 183
Antigen-specific immune complex testing……Page 186
Other tests……Page 187
Further reading……Page 188
Introduction……Page 189
General principles of laboratory control of chemotherapy……Page 191
Sensitivity testing……Page 192
Automation……Page 193
Hospital control and clinical liaison……Page 194
Further reading……Page 195
SECTION 4 MICROBIOLOGY – VIROLOGY, MYCOLOGY AND PARASITOLOGY……Page 197
Histopathology: inclusion bodies and multinucleate giant cells……Page 199
Virology……Page 200
Immunofluorescence……Page 201
Microscopic examination of cell cultures……Page 202
Further reading……Page 204
Methods……Page 205
Uses of electron microscopy……Page 206
Rapid emergency response……Page 211
Conclusions……Page 212
Further reading……Page 213
Choice of cell cultures……Page 215
Preparation of cell cultures……Page 216
Identification of virus growth……Page 217
Further reading……Page 220
Principles……Page 221
Techniques……Page 222
Interpretation and use of serological tests……Page 227
Future of serology……Page 228
Further reading……Page 229
Introduction……Page 231
How automation works in the diagnostic laboratory……Page 233
Should the Laboratory Automate?……Page 234
Further reading……Page 236
Nucleic acid amplification techniques……Page 237
Sequence analysis……Page 244
Further reading……Page 246
Detection of prion infection……Page 249
Diagnosis……Page 251
Antiviral susceptibility tests……Page 252
Cell-mediated immunity……Page 255
Further reading……Page 256
Introduction……Page 257
Microscopic examination……Page 258
Culture……Page 259
Identification……Page 260
Serological tests……Page 261
Further reading……Page 262
Microscopy……Page 263
Films and smears……Page 264
Culture……Page 265
Xenodiagnosis……Page 266
Molecular methods……Page 267
Further reading……Page 268
SECTION 5 HAEMATOLOGY……Page 269
Peripheral blood morphology……Page 271
Examination of bone marrow and bone marrow morphology……Page 273
Further reading……Page 287
Red cell measurements……Page 289
Nucleated RBCs……Page 291
White cell measurements……Page 292
Further reading……Page 295
Screening techniques……Page 297
Detection, identification and quantitation of haemglobinopathies……Page 298
Further reading……Page 303
Introduction……Page 305
Phenotypic tests……Page 306
Genotypic tests for mutation identification……Page 309
Further reading……Page 311
Structure and function……Page 313
Platelet counting……Page 314
Investigations of platelet aggregation……Page 315
Flow cytometry……Page 316
Further reading……Page 317
Blood group identification……Page 319
Methods for blood grouping……Page 320
Antibody screening……Page 321
Transfusion practice……Page 324
Further reading……Page 325
Flow cytometry and immunophenotyping……Page 327
Molecular biology in haematology……Page 331
Summary……Page 334
Further reading……Page 335
Cobalamin (vitamin B(12)) and folate……Page 337
Iron……Page 341
Summary……Page 345
Further reading……Page 346
Clinical features……Page 347
Haemolytic disorders……Page 350
Further reading……Page 356
Prothrombin time……Page 357
Activated partial thromboplastin time (APTT)……Page 359
Thrombin time……Page 361
Fibrinogen……Page 362
Factor assays……Page 363
Automated coagulation……Page 364
Further reading……Page 365
SECTION 6 CYTOGENETICS……Page 367
Chromosome banding……Page 369
Chromosome analysis……Page 376
Further reading……Page 379
Labelling of probes……Page 381
Applications in basic research……Page 382
Applications in cancer diagnosis and research……Page 384
Further reading……Page 386
SECTION 7 CLINICAL CHEMISTRY……Page 387
Introduction……Page 389
Sample collection and analysis……Page 390
The interpretation of laboratory data……Page 391
Further reading……Page 395
Assay design……Page 397
Assay conditions and calibrators……Page 398
Labels……Page 399
Separation methods……Page 400
Accuracy and precision……Page 401
Quality control……Page 402
Specificity, cross-reactivity and interference……Page 403
Further reading……Page 404
Theory……Page 405
Specificity—how electrodes are made selective……Page 408
Electrochemical reactions……Page 410
Glucose electrodes……Page 413
Complex electrodes……Page 414
Other complex enzyme-mediated electrodes……Page 415
Point of care testing (POCT)……Page 416
Safeguards……Page 417
Further reading……Page 418
Light absorption techniques……Page 419
Turbidimetry and nephelometry……Page 421
Luminescence……Page 423
Further reading……Page 427
Principles……Page 429
Instrumentation……Page 431
Applications……Page 437
Conclusions……Page 438
References……Page 439
Introduction……Page 441
Principles of Methodology……Page 442
Specific Examples……Page 443
Quality Control of Dry Reagent Chemistry……Page 447
Summary……Page 450
Further reading……Page 451
Thin-layer chromatography (TLC)……Page 453
High-performance liquid chromatography (HPLC)……Page 455
Gas chromatography (GC)……Page 457
Electrophoresis……Page 458
Further reading……Page 460
SECTION 8 IMMUNOLOGY……Page 461
A short historical perspective……Page 463
Qualitative techniques……Page 464
Quantitative methods……Page 467
Further reading……Page 470
Introduction……Page 471
Assay of serum complement levels……Page 473
Further reading……Page 477
Qualitative and quantitative tests……Page 479
Functional studies……Page 483
The future of cellular immunology……Page 486
Further reading……Page 487
Introduction……Page 489
Assays for circulating immune complexes……Page 490
Serum sickness……Page 491
Systemic lupus erythematosus……Page 492
Cryoglobulinaemia……Page 501
Further reading……Page 504
HLA structure and function……Page 505
The mechanics of HLA typing……Page 506
Molecular typing……Page 507
Summary……Page 510
SECTION 9 MOLECULAR PATHOLOGY……Page 511
Arcturus LCM—PixCell™ and AutoPix™……Page 513
Laser microbeam microdissection……Page 516
Further reading……Page 517
Real-time PCR chemistries……Page 519
TaqMan PCR chemistry—the 5´ nuclease assay……Page 520
TaqMan primer and probe design……Page 522
TaqMan real-time detection……Page 523
Housekeeping genes……Page 524
TaqMan PCR applications in pathology……Page 525
References……Page 528
Overview of methodology……Page 529
In-cell PCR technologies: definitions……Page 530
In-cell amplification of DNA……Page 532
Reaction, tissue and detection controls for use with in-cell DNA PCR assays……Page 534
Problems encountered with in-cell PCR amplification……Page 535
Future work with in-cell PCR-based assays……Page 536
References……Page 537
Affymetrix GeneChip technology……Page 539
Applied biosystems expression array system……Page 541
Illumina ‘Array of Arrays’™……Page 543
Data handling and secondary analysis……Page 544
References……Page 545
Background to CGH and array CGH……Page 547
Overview of comparative genomic hybridization……Page 548
Microarrays……Page 549
References……Page 552
The history of DNA sequencing……Page 555
The dideoxy method of DNA sequencing……Page 556
Automated sequencing……Page 558
The human genome project……Page 560
Advancing areas in nucleic acid sequencing……Page 561
Further reading……Page 562
Index……Page 563