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Henry's Clinical Diagnosis and Management by Laboratory Methods E-Book

Henry's Clinical Diagnosis and Management by Laboratory Methods E-Book

Richard A. McPherson | Matthew R. Pincus

(2017)

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Book Details

Abstract

Recognized as the definitive reference in laboratory medicine since 1908, Henry's Clinical Diagnosis continues to offer state-of-the-art guidance on the scientific foundation and clinical application of today's complete range of laboratory tests. Employing a multidisciplinary approach, it presents the newest information available in the field, including new developments in technologies and the automation platforms on which measurements are performed.

  • Provides guidance on error detection, correction, and prevention, as well as cost-effective test selection.
  • Features a full-color layout, illustrations and visual aids, and an organization based on organ system.
  • Features the latest knowledge on cutting-edge technologies of molecular diagnostics and proteomics.
  • Includes a wealth of information on the exciting subject of omics; these extraordinarily complex measurements reflect important changes in the body and have the potential to predict the onset of diseases such as diabetes mellitus.
  • Coverage of today's hottest topics includes advances in transfusion medicine and organ transplantation; molecular diagnostics in microbiology and infectious diseases; point-of-care testing; pharmacogenomics; and the microbiome.
  • Toxicology and Therapeutic Drug Monitoring chapter discusses the necessity of testing for therapeutic drugs that are more frequently being abused by users.

Table of Contents

Section Title Page Action Price
Front Cover cover
Endsheet 2 IFC1
Half title page i
Associate Editors ii
Henry's Clinical Diagnosis and Management by Laboratory Methods iii
Copyright Page iv
Table Of Contents v
Contributors viii
Preface xiv
Acknowledgments xvii
1 The Clinical Laboratory 1
1 General Concepts and Administrative Issues 2
Key Points 2
Leadership and Management 2
Strategic Planning 3
Quality Systems Management 3
Human Resource Management 5
Laboratory Design and Service Models 5
Regulation, Accreditation, and Legislation 6
Safety 7
Biological Hazards 8
Chemical Hazards 9
Ergonomic Hazards 10
References 10
Selected References 10
References 10.e1
2 Optimizing Laboratory Workflow and Performance 11
Key Points 11
Understanding Workflow 11
Data Collection Techniques 11
Sample and Test Mapping 11
Tube Analysis 12
Workstation Analysis 12
Instrument Audit 13
Test Menu 14
Processing Mode and Load Balancing. 14
Interviews 14
Task Mapping 14
Workflow Analysis 14
Workflow Modeling 15
Pneumatic Tube Transport of Specimens 16
Understanding Technology 16
The Role of Technology: Principles and Pitfalls 16
Optimizing Performance 17
References 19
Selected References 19
References 19.e1
3 Preanalysis 20
Key Points 20
Precollection Variables 20
Physiologic Factors 20
Common Interferences 21
In Vivo 21
Tobacco Smoking 21
In Vitro 22
Collection-Associated Variables 22
Special Issues That May Impact Analysis 22
Immunoassays 22
Specimen Matrix Effects 23
Molecular Diagnostics 23
Effects of Drugs 23
Specimen Collection 23
The Test Order 23
Time of Collection 23
Specimen Acceptability and Identification Issues 24
Blood Collection Overview 24
Anticoagulants and Additives 24
Blood Collection Devices 25
Blood Storage and Preservation 26
Importance of Policies and Procedures 26
Blood Collection Techniques 27
Arterial Puncture 27
Arterial Puncture Technique 27
Finger or Heel Skin Puncture 27
Central Venous Access Devices 28
CVA Collection Technique 28
Urine and Other Body Fluids Collection 28
Urine 28
Special Urine Collection Techniques 29
Urine Storage and Preservation 29
Other Body Fluids 29
Cerebrospinal Fluid 29
Synovial Fluid 30
Pleural Fluid, Pericardial Fluid, and Peritoneal Fluid 30
Specimen Transport 30
Specimen Processing 31
Precentrifugation Phase 31
Centrifugation Phase 31
Equipment 31
References 31
Selected References 31
References 32.e1
4 Analysis 33
Key Points 33
Principles of Instrumentation 33
Spectrophotometry 33
Velocity of Propagation 34
Energy of EMR 34
Scattering of Radiation 34
Rayleigh Scattering 35
Tyndall Effect 35
Raman Scattering 35
Beer-Lambert Law 35
Components of a Spectrophotometer 36
Radiant Energy Sources 36
Wavelength Selectors 37
Filters 37
Sample Containers (Cuvets) 39
Photodetectors 39
Photodiode Arrays. 40
Charge-Transfer Devices. 40
Signal Processors and Readout 40
Quality Assurance in Spectrophotometry 40
Types of Photometric Instruments 40
Reflectometry 41
Reflectometers 42
Molecular Luminescence Spectroscopy (Fluorometry) 42
Principle 42
Components of Fluorometers and Spectrophotofluorometers 42
Nephelometry and Turbidimetry 43
Principle 43
Nephelometer 43
Turbidimetry 43
Refractometry 43
Osmometry 43
Principle of Freezing-Point Osmometry 44
Freezing-Point Osmometer 44
Flow Cytometry 44
Instrument Components 44
Electrochemistry 44
Potentiometry 44
Reference Electrodes 44
Ion-Selective Electrode 45
pH Electrode 45
pCO2 Electrode 46
Coulometry 46
Amperometry 46
pO2 Gas-Sensing Electrode 46
Voltammetry 46
Conductance 46
Impedance 46
Electrophoresis and Densitometry 46
Isoelectric Focusing 47
Chromatography 47
Gas Chromatography 48
Liquid Chromatography 49
Mass Spectrometry 49
Atomic Weights (amu and Da) 50
Mass/Charge Ratio 50
Basic Components 50
Ion Source Unit 50
Mass Spectrometer Analyzer Unit 50
Ion Detector 50
Scintillation Counter 51
Capillary Electrophoresis 52
Nuclear Magnetic Resonance Spectroscopy 53
General Analytic Methods and Issues 53
Chemicals 53
Water 54
Types of Water Purity 54
Purification 54
Monitoring Water Purity 54
Once Your System Has Been Installed 54
Measurement of Mass 54
Types of Balances 54
Unequal-Arm Substitution Balances 54
Magnetic Force Restoration Balance 54
Top-Loading Balances 55
Electronic Balances 55
Calibration 55
Handling Weights Used for Testing Accuracy 55
Environmental Concerns for Best Weighing Accuracy 55
Balance Specifications 55
Laboratory Glassware and Plasticware 55
Types of Glassware 55
Types of Plasticware 55
Volumetric Laboratoryware 56
Pipets 56
Class A Designation 56
Types of Pipets 56
Micropipets 56
Pipet Calibration 56
Volumetric Flasks 56
Calibration of Volumetric Glassware 56
Thermometry 56
Types of Thermometers 56
Special Applications of Temperature-Sensing Devices 57
Mercury-Free Laboratories 57
Thermometer Calibration 57
Water Baths 57
Maintenance 57
Quality Control 57
Heating Blocks, Dry-Bath Incubators, and Ovens 57
Mixing 57
Single-Tube Mixers 57
Multiple-Tube Mixers 57
Aqueous Solution 58
Molarity 58
Millimoles 58
Normality 58
Molal 58
Dilutions 58
Acids, Alkalis, and pH 59
References 59
Selected References 59
References 59.e1
5 Analysis 60
Key Points 60
Evolution of Automation 60
Preanalytic and Postanalytic Automation 61
Automated Delivery 61
Automated Specimen Processing 61
Postanalytic Processing 61
Emerging Technologies in Preanalytic and Postanalytic Processing 62
Automated Specimen Inspection 62
Radio-Frequency Identification 62
The Automated Chemistry Analyzer: Core Components 62
Sampling 62
Reagent Handling 62
Mixing and Incubation 63
Detection 63
Data Processing and Real-Time Monitoring 63
The Continuum of Chemistry Automation 63
Workstation 63
Workcell 63
Total Laboratory Automation 63
Laboratory Automation in Other Sections of the Lab 64
Hematology 64
Microbiology 64
Planning for Laboratory Automation 64
References 65
Selected References 65
References 65.e1
6 Point-of-Care Testing and Physician Office Laboratories 66
Key Points 66
Introduction: Definition of Point-of-Care Testing 66
Regulatory Requirements 66
Laboratory Directorship 67
Compliance 68
Types of Point-of-Care Testing 68
Hematology 68
Chemistry 68
Microbiology 69
Advantages and Disadvantages of Point-of-Care Testing 69
Connectivity and Data Management 69
Implementation of a Point-of-Care Testing Program 70
Physician Offices 70
Hospitals and Medical Centers 70
Point-of-Care Committee and Management Team 70
Selection of Appropriate Tests 71
Selection of Appropriate Testing Devices and Methods 71
Policies and Procedures 71
Method Validations 71
Oversight of Training and Competency 71
Maintenance of a Point-of-Care Testing Program 72
Quality Assurance Program 72
Conclusions 72
References 72
Selected References 72
References 72.e1
7 Postanalysis 73
Key Points 73
Assessment of Analytic Correctness of Results 73
Alarms and Flags 73
Flags for Problem Specimens 74
Flags for Specimens That Require Additional Analysis with Another Method 74
Flags for Problematic Results 74
Delta Checks 74
Assessment of Clinical Significance of Results 74
Critical Values 74
Reference Ranges 74
Definition of Reference Intervals 74
Factors That Influence Reference Ranges 75
Determination of Reference Ranges 75
Variability of Laboratory Results 75
General Principles for the Interpretation of Laboratory Results 75
Diagnostic Accuracy 75
Truth Table 75
Sensitivity and Specificity 75
Effect of Altering the Test Cutoff 77
The Need for High Sensitivity versus High Specificity 77
Predictive Value and Prevalence of Disease 77
Predictive Value and Accuracy 78
Bayes Theorem 78
Likelihood Ratio 79
Receiver Operator Characteristic Curves 80
Positivity Criterion 80
Evidence-Based Medicine 81
Systematic Reviews of Clinical Effectiveness Research and Clinical Practice Guidelines 82
Systematic Reviews 82
Clinical Practice Guidelines 82
References 83
Selected References 83
References 83.e1
8 Interpreting Laboratory Results 84
Key Points 84
Interpreting and Correlating Abnormal Laboratory Values 84
General Considerations 84
Fundamental Principles in Interpretation of Values 84
Abnormalities in the Hematology Profile 85
Anemias 85
Microcytic Anemia 85
Serum Ferritin Levels 86
Use of Serum Iron and Iron-Binding Capacity 86
Use of Red Blood Cell Distribution Width 86
Normocytic Anemia 86
Hyperproliferative Normocytic Anemias 86
Hemolytic Anemia 86
Microangiopathic Hemolytic Anemia 87
Hypoproliferative Normocytic Anemias 87
Bone Marrow Hypoplasia/Aplastic Anemia 87
Myelodysplastic Syndrome 87
Anemia of Renal Failure 88
Macrocytic Anemia 88
Quantitative White Blood Cell Abnormalities 88
Infection 88
Elevated WBC Due to Leukemoid Reaction 88
Elevated WBC Due to Chronic Myelogenous Leukemia 89
Elevated WBC Due to Chronic Lymphocytic Leukemia 89
Leukocytosis Due to Acute Leukemias 89
Low White Cell Counts 89
Aplastic Anemia 89
Gram-Negative Sepsis as a Cause of Leukopenia 89
Coagulation Disorders 89
Abnormalities in Clinical Chemistry: Chemical Pathology 90
Electrolyte Abnormalities 90
Hyponatremia 90
Pseudohyponatremia 91
Hypernatremia 91
Hypokalemia 91
Hyperkalemia 92
Renal Disease 92
BUN and Creatinine 92
Calcium and Phosphate 92
Calcium and Albumin 93
Blood Gas Abnormalities 93
Anion Gap 93
Oxygenation 94
Glucose Abnormalities 94
Other Abnormal Laboratory Findings in Diabetes Mellitus 95
Liver Function Tests 95
Correlations of Liver Function Test Results with Other Laboratory Findings 96
Cardiac Function Tests 96
Diagnosis of Myocardial Infarction and Acute Coronary Syndrome 96
Diagnosis of Congestive Heart Failure 97
Skeletal Muscle Function 97
Pancreatic Function Tests 97
Markers for Inflammatory Conditions 97
Endocrine Function Testing 97
Principle 97
Thyroid Function 97
Adrenal Function 98
Parathyroid Hormone and Vitamin D 98
Vitamin D 98
Examples of Clinical Cases with Clinicopathological Correlations 99
Case A: Electrolyte Disorder 99
Evaluation 99
Case B: Complex Electrolyte Disorder 99
2 Clinical Chemistry 161
14 Evaluation of Renal Function, Water, Electrolytes, and Acid-Base Balance 162
Key Points 162
Introduction 162
Volume and Osmolality of Body Fluid 163
Measurement of Body Fluid Volumes 163
Body Fluid Volumes 163
Composition of the Body Fluid 164
Extracellular Composition 164
Intracellular Composition 164
Measurement of Plasma Osmolality 165
Effect of Hyperglycemia on Serum Na 165
Tonicity 166
Osmolality and Specific Gravity 166
Regulation of Extracellular Volume 166
Nonrenal Control of Water and Electrolyte Balance 167
Insensible Loss of Water from the Skin 167
Loss of Respiratory Water 167
Loss of Water in the Gastrointestinal Tract 167
Measurement of Renal Function 167
Concept of Clearance 167
Quick Formulas for the Calculation of Clearance 168
Conversion of Clearance in L/week to mL/min 168
Measurement of Glomerular Filtration Rate 168
Measurement of GFR with Exogenous Substances 168
Measurement of GFR with Endogenous Substances 168
Creatinine as a Measure of Renal Function 168
Creatinine Measurement 169
Cimetidine-Enhanced Creatinine Clearance 169
The Effect of Drugs on Serum Creatinine 169
Formulas to Estimate Creatinine Clearance as an Estimate of GFR 169
Urea as Measure of Renal Function 170
Measurement of Urea 170
Other Measures of GFR 170
Cystatin C 170
Beta-2 Microglobulin 171
Beta Trace Protein 171
Tryptophan Glycoconjugate 171
Urea Clearance and Urea/Creatinine Ratio in Serum 171
Glomerular Filtration Rate (GFR), Renal Plasma Flow (RPF), and Filtration Fraction (FF) 171
Fractional Excretion (FE) 172
Renal Failure Index (RFI) 172
Fractional Reabsorption (FR) 172
Free Water Clearance and Negative Free Water Clearance 172
Biomarkers of Acute Kidney Injury (AKI) 172
Kidney Injury Molecule-1 (KIM-1) 173
Neutrophil Gelatinase-Associated Lipocalin (NGAL) 173
Interleukin-18 (IL-18) 173
Fatty Acid–Binding Protein (FABP) 173
Disorders of Potassium 174
Control of Transcellular Flux of Potassium 174
Control of Renal Excretion of Potassium 174
Plasma Renin Activity (PRA), Plasma Aldosterone Concentration (PA), and Abnormalities in Potassium Metabolism 175
Causes and Pathogenesis of Hypokalemia 175
Differential Diagnosis of Hypokalemia 176
Causes and Pathogenesis of Hyperkalemia 176
Differential Diagnosis of Hyperkalemia 177
Disorders of Water, Sodium, and Antidiuretic Hormone (ADH) Metabolism 178
Regulation of Thirst and ADH Release 178
Urine Concentration and Dilution 178
Polyuria 179
Osmotic Diuresis 179
Water Diuresis 179
Causes and Pathogenesis of Hyponatremia 180
Causes and Pathogenesis of Hypernatremia 181
Acid-Base Disorders 181
Bicarbonate and CO2 Buffer System 181
Definitions of Acid and Base 181
Whole-Body Acid-Base Balance 181
Net Acid Production 182
Net Acid Excretion 182
Metabolic Acidosis 182
Classification 182
Renal Acidosis 182
Organic Acidosis 183
Lactic Acidosis 183
d-Lactic Acidosis 183
Ketoacidosis 183
Serum Anion Gap (AG) 184
Differential Diagnosis 184
3 Urine and Other Bodily Fluids 441
28 Basic Examination of Urine 442
Key Points 442
Urine Formation 442
Components of Basic (Routine) Urinalysis 443
Specimen Evaluation 443
Gross/Physical Examination 443
Appearance 443
Color 443
Red Urine. 443
Yellow-Brown or Green-Brown Urine. 444
Orange-Red or Orange-Brown Urine. 444
Dark Brown or Black Urine. 444
Blue, Green, or Blue-Green Urine. 444
Clarity (Character) 444
Chyluria. 445
Lipiduria. 445
Odor 445
Urine Volume 445
Increases in Urine Volume 445
Defective Hormonal Regulation of Volume Homeostasis. 445
Defective Renal Salt/Water Absorption. 445
Osmotic Diuresis. 445
Decreases in Urine Volume 445
Prerenal. 445
Postrenal. 445
Renal Parenchymal Disease. 445
Specific Gravity and Osmolality 445
Specific Gravity 446
4 Hematology and Transfusion Medicine 509
30 Basic Examination of Blood and Bone Marrow 510
Key Points 510
Hematology Principles and Procedures 510
Hemoglobin 510
Hemoglobin Derivatives 510
Hemiglobin (Methemoglobin) 510
Sulfhemoglobin 511
Carboxyhemoglobin 511
Measurement of Hb Concentration (Manual Technique of Hemoglobinometry) 511
Hemiglobincyanide Method 511
Principle. 511
Reagent. 511
Method. 512
Errors in Hemoglobinometry 512
Errors Inherent in the Sample. 512
Errors Inherent in the Method. 512
Errors Inherent in the Equipment. 512
Spectrophotometric Identification of Hemoglobins 512
Hematocrit (Packed Cell Volume) 512
Gross Examination 513
Hematocrit Measurement by Micromethod 513
Equipment 513
Procedure 513
Interpretation of Results 513
Sources of Error 513
Centrifugation. 513
Sample. 513
Other Errors. 513
Erythrocyte Indices 513
Mean Cell Volume 513
Mean Cell Hemoglobin 513
Mean Cell Hemoglobin Concentration 513
Manual Blood Cell Counts 514
Erythrocyte Counts: Manual 514
Semiautomated Methods 514
Reticulocyte Count: Manual 514
Principle 514
Reagent. 514
Controls. 514
Procedure 514
Reference Values 514
Interpretation 514
Sources of Variation 514
Physiologic Variation in Erythrocytes 514
Leukocyte Counts: Manual 515
Specimen Collection 515
Hemocytometer Method 515
Counting Chamber. 516
Diluting Fluid. 516
Procedure 516
Sources of Error. 516
Errors due to the nature of the sample. 516
Operator’s errors. 516
Errors due to equipment. 516
Inherent or field errors. 516
Nucleated red blood cells. 516
Reference Value. 516
Physiologic Variation in Leukocytes 516
Platelet Counts 516
Hemocytometer Method—Phase-Contrast Microscope 517
Specimen. 517
Diluent Solution. 517
Procedure 517
Calculation. 518
Sources of Error. 518
Falsely elevated counts. 518
Falsely low counts. 518
Reticulated Platelets 518
Physiologic Variation in Platelets 519
Instrument Technology 519
Hematology Analyzers 519
Electrical Impedance 519
Radiofrequency Conductivity 519
Light Scattering 519
Cytochemistry 519
Reporting/Flagging 520
Sources of Error 520
Automated Differential Leukocyte Counting 520
Digital Image Processing 520
Automated Reticulocyte Counting 521
Blood Film Examination 521
Examination of Wet Preparations 521
Making and Staining Blood Films 521
Wedge Method 521
Cover Glass Method 521
Spinner Method 521
Blood Stains 521
Wright’s Stain 521
Procedure 522
Staining Problems 522
Excessively blue stain. 522
Excessively pink stain. 522
Other staining problems. 522
Other Stains 522
Reference Method 522
Automated Slide Stainer 522
Review of Peripheral Smear 522
Erythrocytes 522
Color 522
Hemoglobin Content 522
Polychromatophilia 523
Size 523
Shape 524
Structure 525
Basophilic Stippling (Punctate Basophilia) 525
Howell-Jolly Bodies 526
Cabot Rings 526
Malarial Stippling 526
Rouleaux Formation 526
Nucleated Red Cells 526
Significance of Nucleated Red Cells 527
Leukoerythroblastic Reaction 527
Leukocytes on Peripheral Blood Smear Examination 527
Leukocytes Normally Present in Blood 527
Neutrophil (Polymorphonuclear Neutrophilic Leukocyte; Segmented Neutrophilic Granulocyte) 527
Eosinophil (Eosinophilic Granulocyte) 528
Basophil (Basophilic Granulocyte) 528
Monocyte 529
Lymphocyte 529
Artifacts 529
Broken Cells 529
Degenerative Changes 530
Contracted Cells 530
Endothelial Cells 530
Radial Segmentation of the Nuclei 531
Vacuolation 531
“Pseudophagocytosis” 531
Sources of Error in the Differential Leukocyte Count 531
Platelets on Peripheral Blood Smear Examination 531
Erythrocyte Sedimentation Rate 531
Plasma Factors 532
Red Cell Factors 532
Stages in the ESR 532
Methods 532
Westergren Method 532
Equipment. 532
Reagent. 532
Procedure 532
Modified Westergren Method 532
Sources of Error 532
Alternative Methods and Technologies to Measure ESR 533
Application 533
Bone Marrow Examination 533
Indications for Marrow Study 533
Preparation of the Aspirate and Biopsy Section 534
Marrow Films 534
Direct Films 534
Imprints 534
Crush Preparations 534
Special Studies 534
Histologic Sections 534
Staining Marrow Preparations 534
Romanowsky’s Stain 534
Perls’ Test for Iron 534
Procedure 534
Interpretation 534
Sections 534
Examination of Marrow 536
Peripheral Blood 536
Cellularity of the Marrow 536
Distribution of Cells 536
Maturation 536
Presence of Rare Cell Types or Abnormal Cells 536
Evaluation of the Biopsy Specimen 538
Interpretation 538
References 538
Selected References 539
References 539.e1
31 Hematopoiesis 540
Key Points 540
Stem Cells 540
Hematopoietic Stem Cells and Progenitors 540
Hematopoietic Growth Factors 541
Hematopoietic Tissues 542
Embryonic and Fetal Hematopoiesis 542
Postnatal Hematopoiesis 542
Erythrocyte Production 542
Normoblastic Maturation 542
Megaloblastic Maturation 544
Regulation of Erythrocyte Production 544
Synthesis of Hemoglobin 544
Heme Synthesis 544
Globin Synthesis 544
Structure and Function of Hemoglobin 546
Erythrocyte Destruction 546
Degradation of Hemoglobin 546
Erythrokinetics 546
Measurements of Total Production of Erythrocytes or Hemoglobin 547
Measurements of Total Destruction of Erythrocytes or Hemoglobin 547
Measurements of Effective Production of Erythrocytes: Reticulocyte Count 547
Measurements of Effective Survival of Erythrocytes in Blood 547
Summary 547
Neutrophils 547
Morphology of Neutrophil Precursors 547
Distribution and Kinetics 550
Function 550
Eosinophils 550
Morphology of Eosinophil Precursors 550
Distribution and Kinetics 551
Function 551
Basophils and Mast Cells 551
Morphology 551
Distribution and Kinetics 552
Function 552
Monocytes and Macrophages 552
Morphology 552
Distribution and Kinetics 553
Function 553
Megakaryocytes 553
Morphology 553
Megakaryocytes in Blood 553
Distribution and Kinetics 554
Function 554
Lymphocytes 554
Primary Lymphoid Tissue 554
B Cell Development: Bone Marrow 554
T Cell Development: Thymus 556
Natural Killer Cells: Bone Marrow 556
Secondary Lymphoid Tissue 556
Lymphocyte Function and Physiology 556
References 557
Selected References 558
References 558.e1
32 Erythrocytic Disorders 559
Key Points 559
Anemias 559
General Manifestations 559
Impaired Production—Iron Deficiency Anemia 559
Iron Metabolism (Beutler, 2010) 559
Iron Deficiency Anemia (IDA) 560
Clinical Features 561
Laboratory Features 561
Blood. 561
Marrow. 561
Serum Iron. 562
Serum (Total) Iron-Binding Capacity. 562
Percent Saturation of TIBC. 562
Serum Ferritin. 562
Erythrocyte Porphyrins. 562
Serum Transferrin Receptors. 562
Serum Transferrin Receptor–to–Serum Ferritin Ratio. 562
Reticulocyte Hemoglobin Content. 562
Hepcidin Level. 562
Differential Diagnosis 562
Management 563
Iron-Refractory Iron Deficiency Anemia (IRIDA) 563
Impaired Production— Megaloblastic Anemia 563
Macrocytosis with Normoblastic Marrow 563
Megaloblastic Anemia 563
Blood 563
Marrow. 563
Erythrokinetics. 564
Cobalamin (Vitamin B12) Metabolism 564
Cobalamin Deficiency 564
Inadequate Intake 564
Defective Production of Intrinsic Factor 564
Pernicious Anemia 564
Clinical Features 564
Gastric Findings 564
Immune Abnormalities 565
Pernicious Anemia in Children 565
Gastrectomy 565
Defective Absorption of Cobalamin 565
Malabsorption Syndromes. 565
Lack of Availability of Cobalamin. 565
Diagnosis of Cobalamin Deficiency 565
Therapeutic Trial. 565
Serum Cobalamin Assay. 565
Methylmalonic Acid and Homocysteine Assays. 565
Deoxyuridine Suppression Test. 565
Detecting the Cause of Cobalamin Deficiency 566
Folic Acid Metabolism 566
The Folate–Cobalamin Relationship 566
Folic Acid Deficiency (Green, 2010) 566
Inadequate Intake of Folate 566
Evolution of Laboratory Abnormalities. 566
Nutritional Folate Deficiency. 566
Liver Disease. 566
Defective Absorption of Folate 566
Increased Requirement for Folate 567
Inadequate Utilization of Folate 567
Diagnosis of Folate Deficiency 567
Serum and Red Cell Folate. 567
Urinary Formiminoglutamic Acid (FIGLU) 567
Deoxyuridine Suppression Test. 567
Plasma Homocysteine Assay. 567
Acute Megaloblastic Anemia 567
Therapy for Megaloblastic Anemia 567
Other Defects of Nucleoprotein Synthesis 567
Congenital Defects 568
Synthetic Inhibitors 568
Refractory Anemias 568
Impaired Production—Other 568
Anemia of Chronic Disease (Means, 2003; Nemeth & Ganz, 2014) 568
Anemia of Renal Insufficiency 568
Anemia in Liver Disease 568
Anemia in Endocrine Disease 569
Anemia Associated with Bone Marrow Infiltration (Myelophthisic Anemia) 569
Aplastic Anemia 569
Clinical Features 569
Etiology 569
Pathogenesis 569
Prognosis 569
Management 570
Idiopathic Acquired Aplastic Anemia 570
Blood 570
Bone Marrow. 570
Erythrokinetics. 570
Acquired Aplastic Anemia Associated with Chemical or Physical Agents 570
Toxic Aplastic Anemias. 570
Ionizing Radiation. 570
Hypersensitive Aplastic Anemias. 570
Acquired Aplastic Anemia Associated with Other Disease 570
Infection. 570
Paroxysmal Nocturnal Hemoglobinuria. 571
Pregnancy. 571
Thymoma. 571
Immunologic Diseases. 571
Inherited Aplastic Anemia 571
Fanconi’s Anemia. 571
Other Inherited Aplastic Anemias. 571
Pure Red Cell Aplasia 571
Transitory Arrest of Erythropoiesis (Transient Aplastic Crises) 571
Transient Erythroblastopenia of Childhood 571
Congenital Red Cell Aplasia (Diamond-Blackfan Anemia; Congenital Hypoplastic Anemia) 571
Acquired Pure Red Cell Aplasia 572
Sideroblastic Anemia 572
Hereditary Sideroblastic Anemias 572
Acquired Sideroblastic Anemias 572
Refractory Anemia with Ring Sideroblasts. 572
Secondary (Drug- or Toxin-Induced) Sideroblastic Anemia 572
Refractory Anemia 572
Congenital Dyserythropoietic Anemias 572
Blood Loss Anemia 573
Acute Posthemorrhagic Anemia 573
Chronic Posthemorrhagic Anemia 573
Hemolysis—General 573
Erythrocyte Survival Studies 573
Hemoglobin Destruction 573
Blood Film 574
Bone Marrow 574
Hemolysis—Membrane Disorders 575
Hereditary Spherocytosis 575
Osmotic Fragility Test 575
Autohemolysis Test 575
Additional Tests 575
Hereditary Elliptocytosis 576
Common HE 576
Hereditary Pyropoikilocytosis 576
Spherocytic HE 576
Southeast Asian Ovalocytosis 576
Hereditary Stomatocytosis 576
Paroxysmal Nocturnal Hemoglobinuria 576
Laboratory Diagnosis 576
Hemolysis—Hemoglobin Disorders 577
Normal Hemoglobins 577
Hb A (α2β2) 577
Hb F (α2γ2) 577
Hb A2 (α2δ2) 577
Embryonic Hemoglobins. 577
The Globin Gene Clusters. 577
Glycosylated Hemoglobins. 577
Laboratory Investigation of Hemoglobinopathies and Thalassemias 578
Cation-Exchange HPLC and Capillary Electrophoresis. 578
Hemoglobin Electrophoresis and Isoelectric Focusing. 578
Acid Elution Slide Test for F Cells. 578
Detection of F cells by Flow Cytometry. 578
Hb A2 Quantitation. 578
Sickling Test—Metabisulfite Slide Test. 578
Sickle Solubility Test. 578
DNA Analysis. 578
Structural Hemoglobin Variants 578
Abnormal Hemoglobin Syndromes 579
Sickling Disorders 581
Sickle Cell Trait (Hb AS) 581
Sickle Cell Disease (Hb SS) 582
Complications. 582
Diagnosis. 584
Hemoglobin SC Disease 584
Hb S/β-Thalassemia 584
Hb SS/α-Thalassemia 584
Hemoglobin SD Disease (Hb S/D–Los Angeles). 584
Hb S/O Arab. 584
Other Compound Heterozygosities with Hb S. 584
Other Common β-Chain Variants 584
Hb C Trait (β6 glu→lys). 584
Hb C Disease. 585
Hb C/β+-Thalassemia. 585
Hb C/β0-Thalassemia. 585
Hb E (β 26 glu→lys). 585
Hb E Trait (Hb AE). 585
Hemoglobin E Disease. 585
Hb E/β-Thalassemia. 585
Hb D Los Angeles (Punjab) (β121 glu→gln). 585
Common α-Chain Variants 585
Hb G Philadelphia (α 68asn→lys). 585
Disorders of Hemoglobin Function and Stability 586
Hemoglobins Associated with High Oxygen Affinity and Polycythemia 586
Hemoglobins Associated with Low Oxygen Affinity 586
M Hemoglobins: Pseudocyanosis 586
Unstable Hemoglobins 586
Heat Instability Test. 586
Isopropanol Precipitation Test. 586
Thalassemias 586
Molecular Defects 587
β-Thalassemias 587
Homozygous β-Thalassemia (Thalassemia Major; Cooley’s Anemia) 587
Heterozygous β-Thalassemia (β-Thalassemia Trait; Thalassemia Minor; Cooley’s Trait) 588
β-Thalassemia Trait with Normal Hb A2 589
δβ0-Thalassemia 589
δβ+-Thalassemia: Lepore Hemoglobins 589
Hereditary Persistence of Fetal Hemoglobin 589
Deletional Pancellular HPFH 589
Hb Kenya 589
Nondeletional Pancellular HPFH 589
Heterocellular or Swiss-type HPFH 589
α-Thalassemias 590
α-Thalassemia Syndromes 590
Hemoglobin Bart’s Hydrops Fetalis (− −/− −). 590
Hemoglobin H Disease (−α/− −). 590
Hemoglobin H preparation. 590
α-Thalassemia Trait: Heterozygous α0-Thalassemia (− −/αα) or Homozygous α+-Thalassemia (−α/−α). 590
Silent Carrier α-Thalassemia (Heterozygous α+-Thalassemia) (αα/−α). 591
Hemoglobin Constant Spring (αCSα/). 591
Screening and Prenatal Diagnosis of Hemoglobin Disorders 591
Hemolysis—Metabolic Disorders 591
Erythrocyte Metabolism 591
Glucose-6-Phosphate Dehydrogenase Deficiency 591
Heinz Bodies 593
Ascorbate Cyanide Test 593
Fluorescent Spot Test 593
Quantitative Assay of G6PD. 593
Pyruvate Kinase (PK) Deficiency 593
Fluorescent Spot Test. 594
Quantitative Assay of PK. 594
Other Glycolytic Enzyme Deficiencies 594
Pyrimidine-5′-Nucleotidase Deficiency 594
Hemolysis—Acquired; Extrinsic 594
Chemical Agents 594
Agents Hemolytic to Normal Cells 594
Agents Hemolytic to Abnormal Cells 594
Physical Agents 594
Heat 594
Traumatic Hemolysis 594
Macroangiopathic Hemolytic Anemia (Cardiac Valvular Disease and Prostheses). 594
Thrombotic Microangiopathy (Microangiopathic Hemolytic Anemia). 595
Hemolytic-Uremic Syndrome 595
Thrombotic Thrombocytopenic Purpura (TTP) 595
Preeclampsia/Eclampsia 595
Infectious Agents 595
Immune Hemolytic Anemias 596
Autoimmune Hemolytic Anemia 596
Etiology and Pathophysiology. 596
AIHA Associated with Warm Antibody. 596
AIHA Associated with Cold Antibody. 597
Cold Agglutinin Disease 597
Paroxysmal Cold Hemoglobinuria 597
AIHA Associated with Warm and Cold Antibodies 597
Alloimmune Hemolytic Disease of the Newborn 597
Drug-Induced Immune Hemolytic Anemia 598
Formation of Ternary Complexes (Neoantigen Formation). 598
Adsorption of Drug to Red Cell Membrane. 598
Induction of Autoantibody by Drugs. 598
Nonimmunologic Adsorption of Immunoglobulins to Red Cell Membrane. 598
Laboratory Investigations of Anemia 599
Macrocytic Anemia (Increased MCV) 599
Megaloblastic Marrow 599
Nonmegaloblastic Marrow 599
Microcytic and Hypochromic Anemias (Decreased MCV and MCH) 599
Normocytic and Normochromic Anemias (Normal MCV) 600
Optimal Marrow Response: Reticulocyte Production Index Greater Than Two 600
Direct Antiglobulin (Coombs’) Test. 600
Inadequate Marrow Response: Reticulocyte Production Index Less Than Two 600
Polycythemia 600
Relative Polycythemia 601
Absolute Polycythemia 601
Appropriately Increased Erythropoietin Production Due to Hypoxia 601
Arterial Oxygen Unsaturation 601
High Oxygen Affinity Hemoglobinopathy 601
Inappropriate Erythropoietin Production 601
Neoplasms 601
Renal Disorders 601
Familial Polycythemia 601
Polycythemia Vera 601
Measurement of Erythrocyte and Plasma Volume 601
Erythrocyte Volume 601
Plasma Volume 602
Interpretation 602
The Porphyrias 602
Physiology 602
Approach to the Porphyrias 602
Clinical Porphyrias and Their Biochemical Basis 602
Neurovisceral Symptoms in Isolation 602
Acute Intermittent Porphyria (AIP) 602
Aminolevulinic Acid Dehydratase Deficiency Porphyria (ADP) 604
Neurovisceral Symptoms Plus Photosensitivity 604
Variegate Porphyria (VP) 604
Hereditary Coproporphyria (HCP) 604
Photosensitivity Plus Hemolytic Anemia 604
Porphyria Cutanea Tarda (PCT) 604
Congenital Erythropoietic Porphyria (CEP) 604
Photosensitivity in Isolation 604
Erythropoietic Protoporphyria (EPP) and X-linked Porphyria (XLP) 604
Analytic Techniques 605
References 605
Selected References 605
References 605.e1
33 Leukocytic Disorders 606
Key Points 606
Leukocytes 606
Nonneoplastic Disorders 606
Granulocytic and Monocytic Disorders 606
Neutrophilia 606
Mechanisms 606
Determinants 607
Neutropenia 607
Morphologic Alterations in Neutrophils 608
Toxic Granulation 608
Döhle Inclusion Bodies 608
May-Hegglin Anomaly 608
Alder-Reilly Anomaly 609
Pelger-Huët Anomaly 610
Chédiak-Higashi Syndrome 610
Functional Disorders of Neutrophils 610
Leukocyte Adhesion Deficiency (LAD) Diseases 610
Eosinophilia 611
Basophilia 611
Monocytosis 611
Monocytopenia 612
Lymphocytic and Plasmacytic Disorders 612
Lymphocytes in Normal Individuals 612
Lymphocytosis 612
Lymphocytosis Associated with Viral Infection 612
Pertussis 612
Chronic Lymphocytosis/Persistent Polyclonal B Cell Lymphocytosis 612
Retrovirus-Associated Diseases and Conditions 613
Infectious Mononucleosis and Epstein-Barr Virus Infection 613
Etiology and Pathophysiology. 613
Clinical Features. 614
Complications. 614
Hematologic Features. 614
Serologic Findings in Immunocompetent Patients. 614
Differential Diagnosis. 614
Course. 614
Cytomegalovirus Infection 614
Toxoplasmosis 614
Autoimmune Lymphoproliferative Syndrome 615
Other Nonneoplastic Causes of Lymphadenopathy 615
Lymphocytopenia 615
Acquired Immunodeficiency Syndrome 615
Etiology. 615
Hematologic Features. 615
Plasmacytosis 615
Leukemoid Reactions 616
Congenital Neutrophilia. 616
Neutrophilic Leukemoid Reactions. 616
Eosinophilic Leukemoid Reactions. 616
Leukoerythroblastosis. 616
Lymphocytic Leukemoid Reactions. 616
Neoplastic Disorders Primarily Involving Leukocytes 616
Overview of Hematopoietic Neoplasms 616
Chronic Myeloproliferative Disorders 617
Chronic Myelogenous Leukemia 617
Clinical Features 617
Laboratory Features 617
Blood. 617
5 Hemostasis and Thrombosis 793
39 Coagulation and Fibrinolysis 794
Key Points 794
Overview of Coagulation and Fibrinolysis 794
Physiologic Hemostasis 795
Endothelium and Platelets 795
Coagulation Protein System 796
Characterization of Coagulation Proteins 796
Physiologic Protein Assemblies 796
The Formation of Fibrin and the Fibrinolytic System 797
The Anticoagulation Protein Systems 798
Protein C/Protein S System 798
Antithrombin 799
Tissue Factor Pathway Inhibitor 799
Prostacyclins and Nitric Oxide and Thrombospondin-5 799
Current Hypothesis for Initiation of the Hemostatic System 799
Clinical Laboratory Hemostasis 800
Physiologic Hemostasis Versus Clinical Assays 800
Screening Tests for Coagulation Disorders 800
Activated Partial Thromboplastin Time 800
Prothrombin Time 801
Thrombin Time 801
Assays Used in Clinical Coagulation Testing 801
Practical Approach to Patients with Coagulation Disorders 801
Global Hemostasis Assays 804
Thrombin Generation 804
Overall Hemostasis Potential 804
Hereditary Coagulation Protein Defects 804
Deficiency of Factor VIII (Hemophilia a) or Factor IX (Hemophilia B) 804
Evaluation of Carriers 805
Treatment of Hemophilia 805
Complications of Treatment 806
Hereditary Deficiencies of Other Coagulation Factors 806
Disorders with Prolonged APTT and Normal PT 807
Factor XI 807
FXII, Prekallikrein, and High Molecular Weight Kininogen 807
Disorders with Prolonged APTT and PT 807
Disorders of Fibrinogen 807
FII Deficiency 808
FV Deficiency 808
FX Deficiency 808
Combined Deficiency of FV and FVIII 808
Combined Deficiency of Vitamin K–Dependent Clotting Factors 808
Disorders with Normal APTT and Prolonged PT 809
FVII Deficiency 809
Disorders with Normal APTT and PT 809
FXIII Deficiency 809
Hereditary Hemorrhagic Disorders of Fibrinolysis 809
Acquired Coagulation Disorders 809
Thrombotic Disorders and Disseminated Intravascular Coagulation 809
Liver Disease 810
Vitamin K Deficiency 810
Massive Transfusion 810
Acquired Coagulation Protein Inhibitors and Lupus Anticoagulant 810
References 811
Selected References 811
References 811.e1
40 Platelet Disorders and von Willebrand Disease 812
Key Points 812
Normal Platelet Biology 812
Platelet Structure 812
Platelet Membrane Glycoproteins and Phospholipids 813
Role of Platelets in Hemostasis and Platelet-Activation Mechanisms 813
Platelet Activation and Signaling Events (see Fig. 40-2) 813
Platelet Coagulant Activity 814
Laboratory Evaluation of Platelet Function Disorders 814
Initial Laboratory Evaluation 814
Studies of Platelet Function 814
Bleeding Time 814
PFA-100/200 816
Platelet Aggregation and Secretion Studies 816
Evaluation of Platelet Function Using Flow Cytometry 816
Quantitative Platelet Disorders 816
Thrombocytopenia 816
Congenital Thrombocytopenias 817
Autosomal Dominant Thrombocytopenias 817
Autosomal Recessive Thrombocytopenias 819
Sex-Linked Inherited Thrombocytopenias 819
Immune Thrombocytopenias 819
Drug-Induced Immune Thrombocytopenia (DITP) 820
Thrombocytosis 821
Inherited Disorders of Platelet Function 821
Overview 821
Disorders of Platelet Adhesion 822
Bernard-Soulier Syndrome 822
Disorders of Platelet Aggregation 823
Glanzmann Thrombasthenia 823
Disorders of Platelet Secretion and Granules 823
Deficiency of Granule Stores 823
δ-Storage Pool Deficiency 823
Gray Platelet Syndrome 824
Quebec Platelet Syndrome 824
Disorders of Platelet Secretion and Signaling Pathways 824
Defects in Receptors 824
Defects in G-Proteins and Their Activation 824
Defects in Phospholipase C Activation, Calcium Mobilization, and Protein Phosphorylation 825
Abnormalities in Arachidonic Acid Metabolism and Thromboxane Synthesis 825
Defects in Cytoskeletal Linking Proteins 825
Wiskott-Aldrich Syndrome 825
Kindlin-3 Deficiency–Leukocyte Adhesion Deficiency 825
Disorders of Platelet Procoagulant Activities: Scott Syndrome 825
Transcription Factor Mutations and Associated Platelet Dysfunction 825
RUNX1/CBFA2 Mutations (Familial Platelet Disorder with Predisposition to Acute Myelogenous Leukemia) 825
GATA-1 Mutations 825
FLI1 825
GFI1B 826
Relative Frequencies of Various Inherited Platelet Disorders 826
Evaluation of Patients with Suspected Congenital Platelet Disorders 826
Therapy of Congenital Platelet Function Defects 826
Acquired Disorders of Platelet Function 826
Myeloproliferative Neoplasms 826
Acute Leukemias and Myelodysplastic Syndromes 827
Dysproteinemias 827
Uremia 827
Acquired Storage Pool Disease 827
Antiplatelet Antibodies and Platelet Function 828
Drugs That Inhibit Platelet Function 828
Von Willebrand Factor, von Willebrand Disease, and Thrombotic Thrombocytopenic Purpura 829
Von Willebrand Factor Biology 829
Laboratory Tests for von Willebrand Factor 830
Von Willebrand Disease 830
Type 1 vWD: Partial Quantitative Abnormality 830
Type 3 vWD: Severe Quantitative Abnormality 831
Qualitative Abnormalities: Type 2A, Type 2B, Type 2M, and Type 2N vWD 831
Type 2A 831
Type 2B 831
Type 2M 831
Type 2N (Normandy) 831
Platelet-Type vWD 831
Acquired von Willebrand Syndrome 831
Thrombotic Thrombocytopenic Purpura 832
References 833
Selected References 833
References 833.e1
41 Laboratory Approach to Thrombotic Risk 834
Key Points 834
Endogenous Anticoagulant Pathways 834
Thrombophilic Proteins or Factors 835
Antithrombin 835
Protein C 837
Protein S 837
Activated Protein C Resistance and Factor VLeiden 838
Prothrombin g20210a 838
Elevation of Other Coagulation Factor Levels 838
Tissue Factor Pathway Inhibitor 838
Dysfibrinogenemia 839
Annexin A5 839
Hypofibrinolytic Mechanisms 839
Acquired Hypercoagulable States 839
Antiphospholipid Syndrome and Lupus Anticoagulant 839
Other Coagulation Abnormalities 840
Hyperhomocysteinemia 841
General Aspects of the Laboratory Evaluation for Thrombotic Risk 841
References 841
Selected References 841
References 841.e1
42 Antithrombotic Therapy 842
Key Points 842
Vitamin K Antagonists 842
Antithrombin-Dependent Parenteral Anticoagulants 845
Unfractionated Heparin 845
Heparin Resistance 846
Low Molecular Weight Heparin 846
Fondaparinux 846
Heparin-Induced Thrombocytopenia 846
Target-Specific Anticoagulants 849
Parenteral Agents 849
Oral Agents 849
Antiplatelet Therapy 851
Aspirin 851
P2Y12 Inhibitors 852
Glycoprotein IIb/IIIa Inhibitors 853
References 853
Selected References 853
References 853.e1
6 Immunology and Immunopathology 855
43 Overview of the Immune System and Immunologic Disorders 856
Key Points 856
Lymphoid Cells 856
T Lymphocytes 856
B Lymphocytes 857
Antigen-Presenting Cells 857
Natural Killer Cells 857
Nonlymphoid Cells 857
Neutrophils and Eosinophils 857
Basophils and Mast Cells 857
Humoral Factors 857
Immunoglobulins 857
Complement 858
Cytokines 858
Histocompatibility Antigens 858
Mechanisms of Immunologic Injury 859
Laboratory Applications of Immunologic Assessment 859
Therapeutics in Immunology 860
References 861
Selected References 861
References 861.e1
44 Immunoassays and Immunochemistry 862
Immunoassays and Immunochemistry 862
General Characteristics of Antigen–Antibody Reaction 862
Characteristics of Antigens 863
Characteristics of Antibodies 863
Polyclonal Antibodies 863
Monoclonal Antibodies 863
Antibody Production by Recombinant Technology 863
Kinetics of Antigen–Antibody Reaction 863
Overview of General Principles of Immunoassays 863
Classes of Immunoassays 863
Conjugation Chemistry 864
Characteristics of the Solid Phase 864
Precipitin and Nephelometric Immunoassays 865
Background and Principles of Precipitin Reaction 865
Nephelometric Immunoassays 866
Particle Immunoassay 866
Principle of Particle Agglutination 866
Hemagglutination 866
Gelatin Particle Agglutination 868
Latex Agglutination 868
Latex Turbidimetric Assay 868
Particle-Counting Immunoassay 868
Other Particle Immunoassays 868
Summary 868
Radioimmunoassay 870
Background 870
Assay Principles and Methods 870
Summary 872
Enzyme Immunoassay 872
Background and Classification 872
Heterogeneous Enzyme Immunoassays 872
Colorimetric Enzyme Immunoassay 873
Fluorescent Enzyme Immunoassay 873
Chemiluminescent Enzyme Immunoassay 873
Homogeneous Enzyme Immunoassays 874
Background 874
Enzyme-Multiplied Immunoassay Technique 874
Substrate-Labeled Fluorescent Immunoassay 875
Apoenzyme Reactivation Immunoassay 875
Enzyme Inhibitory Homogeneous Immunoassay 875
Cloned Enzyme Donor Immunoassay 877
Summary 878
Fluorescent Immunoassay 878
Background and Classification 878
Heterogeneous Fluorescent Immunoassay 878
Fluoroimmunometric Method 878
Radial Partition Immunofluorometric Assay 879
Time-Resolved Fluoroimmunoassay 879
Homogeneous Fluorescent Immunoassay 879
Fluorescence Polarization Assay 879
Fluorescence Excitation Transfer Immunoassay 880
Fluorescent Protection Immunoassay 880
Chemiluminescent Immunoassay 880
Background 880
Chemiluminescent Immunoassay Using Acridinium Esters as Labels 880
Electrochemiluminescent Immunoassay 880
Noncompetitive Immunoassay for Small Molecules 881
Background 881
Anti-Idiotype Antibody-Based Immunoassay 881
Open-Sandwich Immunoassay 882
Antimetatype Antibody–Based Immunoassay 882
Sensitive Immunoassay Using Specimen-Pretreatment Methods 882
Background 882
Highly Sensitive Immunoassay for Viral Antigen Using Specimen-Pretreatment Method 882
Instrument Automation and Modulation of Assay Systems 883
Homogeneous Assay Systems 883
Heterogeneous Immunoassay Systems 883
Practical Flow of the Immunoassay in the Analytic System 883
Instrumentation and Key Points for Heterogeneous Immunoassay 884
Reaction Cuvette 884
Sampling and Fluid Delivery Type 884
Carryover 884
Bound/Fixed Separation and Washing Systems 884
High-Throughput Fully Automated Heterogeneous Immunoassay Systems 884
Remote Maintenance 885
Rapid and Simple Test Devices for Point-of-Care Testing 885
Background 885
Immunochromatographic Devices 885
Magnetic Biosensor Assay 886
Summary 887
Simultaneous Multiple Immunoassays 887
Background 887
Microspot Assay 887
Multianalyte Microarray Immunoassay 887
Flow Cytometric Immunoassay 887
Microchannel Assay Using Compact Discs 888
7 Medical Microbiology 1071
56 Viral Infections 1072
Key Points 1072
Introduction 1072
Cell Culture 1074
Antigen Detection 1074
Nucleic Acid Amplification Testing (NAAT) 1074
Viral Serology 1076
Clinical Viral Infectious Syndromes 1076
Herpetic Mucocutaneous Infections 1077
Specimen Collection and Handling Guidelines 1078
Cell Culture Isolation of Herpes Simplex Virus 1078
Direct Detection of Herpes Simplex Virus 1079
Nucleic Acid–Based Detection of Herpes Simplex 1079
Serologic Diagnosis 1079
Pediatric and Adult Viral Respiratory Syndromes 1079
Influenza 1081
Bronchiolitis 1081
Croup 1081
Metapneumovirus 1082
Adenovirus 1082
Coronavirus 1083
Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) 1083
Middle Eastern Respiratory Syndrome Coronavirus (MERS-CoV) 1083
Bocaviruses 1083
Rhinoviruses (HRV) 1083
Specimen Collection 1083
Virus Antigen Assays 1083
Virus Isolation 1084
Molecular Detection Methods 1084
Infectious Mononucleosis and Related Infections 1084
Diagnosis of Infectious Mononucleosis 1084
Heterophile-Negative Infectious Mononucleosis 1085
Chronic Fatigue Syndrome 1085
Congenital and Perinatal Viral Infections 1085
Cytomegalovirus 1086
Rubella 1087
Herpes Simplex Virus 1087
Human Immunodeficiency Virus, Parvovirus, Enterovirus, and Hepatitis B Virus 1087
Viral Meningitis and Encephalitis 1088
Laboratory Diagnosis 1089
Viral Exanthems and Common Cutaneous Infections 1090
Viral Gastroenteritis 1091
Laboratory Diagnosis 1092
Viral Hepatitis 1092
Hepatitis A Virus (HAV) 1092
Hepatitis E Virus (HEV) 1092
Hepatitis B Virus (HBV) 1092
Hepatitis C Virus (HCV) 1093
Acquired Immunodeficiency Syndrome 1093
Virus-Associated Neoplasia 1097
Human Papilloma Viruses (HPV) 1097
Emerging Infections 1097
Ebola Viral Disease (EVD) 1097
References 1097
Selected References 1098
Websites and E-Mail Addresses 1098
References 1098.e1
57 Chlamydial, Rickettsial, and Mycoplasmal Infections 1099
Key Points 1099
Chlamydial Infections 1099
Structure 1099
Replication 1099
Chlamydia trachomatis 1100
Epidemiology, Pathology, and Clinical Manifestations 1100
Chlamydophila (Formerly Chlamydia) psittaci 1101
8 Molecular Pathology 1299
65 Introduction to Molecular Pathology 1300
Key Points 1300
The Molecular Biology Revolution and Its Impact on the Practice of Pathology 1300
Discussion of Diagnostic Molecular Pathology 1300
Application of Molecular Pathology to Detection of Cancer 1301
Implications of Molecular Diagnostics on the Practice of Pathology and Medicine 1301
Redefining Disease 1301
Use of Molecular Biology in the Treatment of Disease 1301
Data Analysis 1301
Quality Assurance 1302
Some Caveats 1302
References 1302
Selected References 1302
References 1302.e1
66 Molecular Diagnostics 1303
Key Points 1303
Nucleic Acid Biochemistry and Biology 1303
Molecular Composition and Structure 1303
Nucleic Acid–Associated Enzymes 1305
Replication of DNA 1305
Transcription of DNA to RNA 1306
Posttranscriptional Modification 1306
Translation of RNA to Protein 1306
Transcriptional Control 1306
Gene Regulation Mediated by Small RNA 1307
Epigenetics and Gene Regulation 1307
Mechanisms of DNA Repair 1307
DNA Mutations 1308
Nucleic Acid Analyses 1308
Electrophoretic Separation 1309
Nucleic Acid Hybridization 1309
Hybridization Assays: Basic Components 1309
Probe 1310
Sample 1311
Controlled Conditions Permissive for Complementary Base Pairing 1311
Detection of Hybrids 1311
Hybridization Assay Formats 1311
Liquid or Solution Phase Hybridization 1311
Solid-Support Hybridization 1312
Dot or Linear Array Hybridization 1312
Southern and Northern Hybridizations 1312
Microarray Hybridization (“DNA Chip Technology” or “Bead Arrays”) 1312
In Situ Hybridization 1312
Amplification Methods 1313
Polymorphism Detection Assays 1313
DNA Sequencing 1313
Relationship to Laboratory Evaluation of Disease 1313
Molecular Diagnosis 1313
Beyond Diagnosis 1314
References 1314
Selected References 1315
References 1315.e1
67 Polymerase Chain Reaction and Other Nucleic Acid Amplification Technology 1316
Key Points 1316
Target Amplification Methods 1316
Polymerase Chain Reaction 1316
Reverse-Transcriptase Polymerase Chain Reaction 1316
Nested Polymerase Chain Reaction 1317
Multiplex Polymerase Chain Reaction 1317
End-Point Quantitative Polymerase Chain Reaction 1318
Real-Time (Homogeneous, Kinetic) Polymerase Chain Reaction 1319
Rapid-Cycle Polymerase Chain Reaction 1321
Digital Polymerase Chain Reaction 1321
Transcription-Based Amplification 1321
Strand-Displacement Amplification 1322
Loop-Mediated Amplification 1323
Helicase-Dependent Amplification 1324
Nicking Endonuclease Amplification 1324
Probe Amplification Methods 1324
Cleavase/Invader Technology 1324
Signal Amplification Methods 1324
Branched DNA 1326
Hybrid Capture Assays 1326
Summary 1326
References 1326
Selected References 1326
References 1327.e1
68 Hybridization Array Technologies 1328
Key Points 1328
Array Technologies 1328
Macroarrays 1329
Microarrays 1329
Microarray Substrates 1329
Microarray Fabrication 1329
Delivery Technologies 1329
In Situ Synthesis 1330
Oligonucleotide Microarrays 1330
CDNA Microarrays 1331
Sequencing Arrays 1331
Hybrid Capture 1331
Reverse Line-Blot Hybridization 1332
Hybridization, Detection, and Image Analysis 1332
Bioinformatics 1333
Intellectual Property Issues 1334
Clinical Applications of Array Technology 1335
Array Technology in the Clinical Laboratory 1335
Array Technology in Clinical Disease 1335
Limitations 1335
References 1336
Selected References 1336
References 1336.e1
69 Applications of Cytogenetics in Modern Pathology 1337
Key Points 1337
Definitions 1337
Cytogenetics 1338
Chromosomes 1338
Chromosome Structure 1338
Cell Culture 1338
Specimens 1338
Cell Culture Technique 1339
Staining 1340
Karyotype Analysis 1340
Computer-Assisted Imaging 1340
Fluorescence in Situ Hybridization 1341
Technique 1341
Multicolor Fluorescence in Situ Hybridization 1343
Microarray Technology 1343
Chromosome Abnormalities 1344
Numerical Abnormalities 1345
Euploidy 1345
Aneuploidy 1345
Structural Chromosome Abnormalities 1345
Conclusions 1349
Nomenclature 1349
Clinical Applications 1350
CONSTITUTIONAL CYTOGENICS 1350
Prenatal 1350
Postnatal 1351
Childhood and Adult 1352
Cancer Genetics 1352
Cytogenetic Disorders 1353
Chromosomal Aneuploidy Syndromes 1353
Autosomal Aneuploidies 1353
Sex Chromosome Aneuploidies 1354
Other Sex Chromosome Abnormalities 1356
Structural Chromosome Anomalies 1356
Microdeletion Syndromes and Contiguous Gene Syndromes 1356
Other Cytogenetic Phenomena 1358
Fragile X Syndrome 1358
Breakage Syndromes 1358
Summary 1358
References 1358
Selected References 1359
Further Reading 1359
References 1359.e1
70 Establishing a Molecular Diagnostics Laboratory 1360
Key Points 1360
Special Considerations for Molecular Diagnostics Laboratories 1361
Infectious Disease 1361
Cancer 1361
Inherited Disorders 1361
Regulation and Regulatory Agencies 1362
Clinical Laboratory Improvement Acts of 1988 and 2003 1362
Genetic Information Nondiscrimination Act 1362
Food and Drug Administration 1362
Analyte-Specific Reagents 1363
In Vitro Multivariate Index Assay or IVDMIA Draft Guidance 1363
Laboratory Design and Requirements 1363
Laboratory Design 1364
Practices to Aid in Contamination Control 1364
Equipment 1365
Personnel 1365
Laboratory Director 1365
Technical Supervisor 1365
Medical Technologists and Molecular Biology Technicians 1365
Certification of Personnel in Molecular Diagnostics 1366
Financial Management 1366
Reimbursement for Molecular Diagnostics Tests 1366
Patent Issues 1367
Laboratory Information System 1367
Test Management 1368
Menu/Selection 1368
Methods 1368
Clinical Test Formats 1368
Automated Platforms for Molecular Testing 1369
Quality Assurance and Quality Control of the Testing Process 1369
Quality Assurance 1369
Assay Design and Development 1370
New Test Verification/Validation 1371
Analytical Verification 1371
Clinical Verification 1375
Quality Control of the Testing Process 1375
Quality Control of Equipment 1375
Conclusion 1376
Selected References 1376
References 1376.e1
71 Molecular Diagnosis of Genetic Diseases 1377
Key Points 1377
Choice of Techniques 1378
Choice of Applications 1379
Special Concepts Unique to Molecular Genetic Disorders 1381
Molecular Heterogeneity 1381
Variable Penetrance and Expressivity 1381
Uniparental Disomy 1381
Imprinting 1381
Anticipation 1381
Epigenetic Influences and Nonmendelian Inheritance 1382
Allele Frequencies and Mass Population Screening 1382
Predictive Genetic Testing 1382
Specific Disease Examples 1382
Cystic Fibrosis 1382
Duchenne Muscular Dystrophy 1383
Sickle Cell Anemia and Other Hemoglobinopathies 1384
Hereditary Thrombophilias 1384
Trinucleotide Repeat Expansion Disorders 1385
Fragile XA and Fragile XE Syndromes 1385
Neurodegenerative Disorders: Huntington Disease, X-linked Spinal and Bulbar Muscular Atrophy, Spinocerebellar Ataxias, and Dentatorubral-Pallidoluysian Atrophy 1386
Myotonic Dystrophy 1386
Friedreich Ataxia 1386
Laboratory Testing for Trinucleotide Repeat Disorders 1388
Prader-Willi and Angelman Syndromes 1388
Familial Cancers 1388
Tumor Suppressor Genes: Retinoblastoma as a Paradigm 1389
Oncogenes: Multiple Endocrine Neoplasia Type 2 as a Paradigm 1390
Hereditary Breast-Ovarian Cancer 1390
The Hereditary Colorectal Cancer Syndromes 1390
Hereditary Nonpolyposis Colorectal Cancer 1390
Familial Adenomatous Polyposis and Attenuated Familial Adenomatous Polyposis 1391
MYH-Associated Polyposis 1391
Laboratory Testing for Familial Cancer Mutations 1391
Hemochromatosis 1391
Spinal Muscular Atrophy 1392
Mitochondrial DNA Disorders 1392
Other Targets of Molecular Genetic Testing and Screening 1392
The New Frontier of Diagnostic Molecular Genetics: Genome-Level DNA Sequencing 1393
References 1394
Selected References 1394
References 1395.e1
72 Identity Analysis 1396
Key Points 1396
Historical Background 1396
Advantages of DNA 1397
Genetic Markers 1397
Samples and Specimen Collection 1397
DNA Extraction and Quantification 1398
Test Methods and Marker Systems 1398
Short Tandem Repeats 1398
X and Y Chromosome Markers 1400
Mitochondrial DNA Sequencing 1400
Other Systems 1400
Analysis and Use of Test Data 1401
DNA in the Crime Laboratory 1401
DNA in the Pathology Laboratory 1401
Exclusion of Parentage 1401
Probability of Exclusion 1402
Combined Probability of Exclusion 1403
Inclusion of Parentage 1403
Paternity Index Calculation 1403
Likelihood of Parentage 1404
Estimating Parentage with an Absent Parent 1404
Reconstruction of Families 1405
Software Systems 1405
Quality Assurance Standards 1405
Conclusion 1405
References 1406
Selected References 1406
References 1406.e1
73 Pharmacogenomics and Personalized Medicine 1407
Key Points 1407
Overview 1407
Basic Concepts 1407
Drug-Metabolizing Enzyme Pharmacogenomics 1408
Phase I Enzymes 1408
Phase II Enzymes 1410
Uridine Diphosphate Glucuronosyltransferases 1410
Sulfotransferases 1412
Glutathione S-Transferases 1412
N-Acetyltransferase 1412
Thiopurine S-Methyltransferase 1412
Drug-Transporter Pharmacogenomics 1413
ATP-Binding Cassette (ABC) Transporters 1414
ABCB1 1414
ABCC1 and ABCC2 1414
ABCG2 1416
Solute Carrier Transporters 1416
Organic Anion Transporting Polypeptides 1416
Organic Cation Transporters (OCTs) 1417
Organic Anion Transporters (OATs) 1418
Drug Target Pharmacogenomics 1418
DNA Repair Genes 1418
Thymidylate Synthase 1418
Epidermal Growth Factor Receptor 1419
Human Leukocyte Antigen Pharmacogenomics 1423
Pharmacogenomics in Cancer Chemotherapy 1424
5-Fluorouracil 1424
Irinotecan 1425
Tamoxifen 1426
Pharmacogenomics in Other Diseases 1428
Diabetes 1428
Human Immunodeficiency Virus 1428
Osteoporosis 1429
Psychiatric and Cognitive Disorders 1429
Epigenomics 1429
Conclusion 1430
References 1430
Selected References 1430
References 1430.e1
9 Molecular Pathology 1431
74 Diagnosis and Management of Cancer Using Serologic and Other Body Fluid Markers 1432
Key Points 1432
Serum Markers as an Effective Tool for Diagnosis and Monitoring of Cancer 1433
Functional Classification of Tumor Markers 1433
Oncoproteins are Markers for Cell Proliferation 1434
Tumor Suppressors/Cell Differentiation 1434
Adhesion Molecules and Metastasis 1434
Monoclonal Antibody–Defined Tumor Markers 1434
Other Markers 1434
Clinical Applications 1435
Screening 1435
Diagnosis 1435
Prognosis: Recurrence, Metastasis, and Survival 1435
Monitoring Treatment Response 1436
Recommendations for Ordering Tumor Marker Tests 1436
Individual Tumor Markers 1438
α-Fetoprotein 1438
Angiogenic Factors 1438
β2-Microglobulin 1438
Serum Free Light Chain Determinations 1439
Carcinoembryonic Antigen 1440
CA 15-3 and CA 27.29 1440
CA 19-9, CA 50, and CA 19-5 1440
CA 125 1441
CA 72-4 1441
Calcitonin 1441
Chromogranin A 1441
Cytokeratin 19 Fragment 1441
Human Chorionic Gonadotropin 1441
HER2/neu (c-erbB-2) ONCOPROTEIN 1442
p53 1442
Parathyroid Hormone–Related Peptide 1442
Serum Markers for Prostate Cancer 1442
Serum Prostate-Specific Antigen 1442
Methods to Improve the Performance of Serum PSA Measurement for the Early Detection of Prostate Cancer 1443
Free PSA, Complex PSA, and Percentage of Free PSA 1443
proPSA and Prostate Health Index 1444
PSA Doubling Time, Velocity, and Density 1444
Circulating Tumor Cells in Peripheral Blood 1444
Circulating Nucleic Acids in Peripheral Blood 1445
Cell-Free Nucleic Acid Testing in Cancer 1445
Gene Arrays Detecting Oncoprotein Abnormalities 1446
Circulating microRNA 1446
Gene Mutations 1446
Microsatellite Alterations 1446
Promoter Hypermethylation 1446
Cell DNA Testing for Cancer Using Fluorescence in Situ Hybridization 1447
Markers in Other Body Fluids 1447
Urine Markers for Prostate Cancer 1447
Urine Prostate Cancer Antigen 3 1447
Urine TMPRSS2:ERG and Mi-Prostate Score 1447
Prostarix 1447
Urine Hypermethylated Glutathione S-Transferase pi 1 Gene 1448
Urine Metabolic Test for Adenomatous Polyps of Colon 1448
Fecal Occult Blood Testing and Mutant Protein Markers in Stool 1448
Conclusions 1448
References 1448
Selected References 1449
References 1449.e1
75 Oncoproteins and Early Tumor Detection 1450
Key Points 1450
Overview 1450
Cell Biology and Mitogenesis 1450
Signal Transduction Pathways 1451
Oncoproteins in Tumor Detection 1451
Growth Factors 1452
Transforming Growth Factors α and β 1452
Platelet-Derived Growth Factor 1453
Basic Fibroblast Growth Factor 1453
Epidermal Growth Factor and Hepatocyte Growth Factor 1453
Growth Factor Receptors 1454
Epidermal Growth Factor Receptor 1455
HER2/neu Receptor 1455
HER2/neu Detects Pulmonary Neoplasms 1455
HER2/neu in the Detection of Hepatocellular Carcinomas 1455
p185erbB-2 ECD in Other Tumors 1455
E-Cadherin 1456
Metalloproteases in Pancreatic Cancer 1456
G-Proteins 1456
ras-p21 Protein 1456
ras-p21 in Lung and Colon Cancer 1456
ras-p21 in Angiosarcoma 1457
ras-p21 in Pancreatic Cancer 1457
Cytosolic Mitogenic Kinases 1457
Raf 1457
Nuclear Oncoproteins 1457
p53 and c-myc Proteins 1457
Detection of Malignancies by Assaying for p53 Protein 1457
Hepatocellular Carcinoma. 1457
Breast and Lung Cancers. 1457
Colon Cancer. 1458
Bladder Cancer. 1458
Leukemia. 1458
Circulating Anti-p53 Antibodies in Tumor Detection 1458
Negative Regulator of p53, S100, as a Tumor Marker for Malignant Melanoma. 1458
Antibodies to p53 Are Present in the Sera of Many Patients with Epithelial Cell Tumors and Lymphomas. 1458
Anti-p53 in Ovarian Cancer. 1458
Anti-p53 in Hepatocellular Carcinoma. 1458
Anti-p53 in Oral and Esophageal Cancers. 1458
Anti-p53 in Lung Cancers. 1458
Anti-p53 Predicts Cancer Occurrence in Patients with Asbestos Exposure. 1458
Anti-p53 in Colorectal Cancer. 1459
myc Oncogene–Encoded Protein in Tumor Detection 1459
Serum Anti–c-myc Protein Antibodies in Tumor Detection in Breast Cancer, Ovarian Cancer, Leukemias, and Lymphomas 1459
Combined Oncogene Marker Proteins for Detection of Colorectal Cancer 1459
Nuclear Matrix Proteins and Detection of Bladder Cancer 1459
NMP-22 Is an Excellent Biomarker for Bladder and Urothelial Cancers 1459
Bladder Tumor Antigen 1459
Minichromosome Maintenance Proteins 1460
Use of Multiple Oncoprotein Markers in the Diagnosis of Cancer 1460
Multiple Oncoprotein Assay in Patients at Risk for Tumor Development in Patients with Pneumoconiosis 1460
Oncoprotein Arrays Likewise Hold Promise in Detecting Antioncoprotein Antibodies in Sera 1460
Proteomic Approaches to Early Detection of Cancer in Serum 1460
Application of the Proteomic Approach to the Diagnosis of Cancers 1461
Ovarian Cancer 1461
Prostate Cancer 1461
Proteomics Show Promise in Early Tumor Detection 1461
Protein Arrays for Specific Types of Cancer 1461
Squamous Cell Carcinomas of the Head and Neck 1461
Breast Cancer 1461
Lung Cancer 1461
Diagnostic Efficacy of Serum Oncoproteins 1461
How Specific Is Oncoprotein Expression for Different Cancer Types? 1461
Sensitivities of Oncoprotein Expression 1463
Origins of Malignancies 1463
Tumor Size and Oncoprotein Levels 1463
Evaluation and Conclusions 1463
References 1463
Selected References 1463
References 1464.e1
76 Molecular Diagnosis of Hematopoietic Neoplasms 1465
Key Points 1465
Role of Clinical Molecular Diagnostics in Hematologic Cancers 1465
Molecular Diagnosis of Leukemias 1466
Gene Fusion Concept in Leukemia and the Basis for Reverse-Transcription Polymerase Chain Reaction Analysis 1466
Acute Myeloid Leukemias 1467
Acute Promyelocytic Leukemia: t(15;17)(q22;q21)/PML-RARA Abnormality 1468
Core Binding Factor–Related Acute Myeloid Leukemias: t(8;21)(q22;q22)/RUNX1-RUNX1T1 and Inv(16)(p13q22) or t(16;16)(p13;q22)/ CBFB-MYH11 Abnormalities 1470
Acute Myeloid Leukemia with Other Translocations 1470
AML with 11q23/MLL 1470
AML with t(6;9)(p23;q34)/DEK-NUP214 1471
AML with inv(3)(q21q26.2) or t(3;3)(q21;q26.2)/RPN1-EVI1 1471
AML (Megakaryoblastic) with t(1;22)(p13;q13)/RBM15-MKL1 1471
Acute Myeloid Leukemias with FLT3, NPM1, and CEBPA Gene Mutations 1471
FLT3 Mutations in Acute Myeloid Leukemia 1471
NPM1 Mutations in Acute Myeloid Leukemia 1472
CEBPA Mutations in Acute Myeloid Leukemia 1472
Other Gene Mutations in Acute Myeloid Leukemias 1472
WT1 Mutations in Acute Myeloid Leukemias 1472
Acute Lymphoblastic Leukemia/Lymphoma—B and T Cell Lineage 1473
B Cell Lymphoblastic Leukemia/Lymphoma (Precursor B Cell Acute Lymphoblastic Leukemia, B-ALL) 1473
Major Translocation Fusion Gene Abnormalities in B Cell ALL 1473
Prenatal Origins of Childhood Leukemias 1476
BCR-ABL1–Like B-Acute Lymphoblastic Leukemia 1476
T Cell Lymphoblastic Leukemia/Lymphoma 1476
Molecular Diagnosis of Myeloproliferative Neoplasms 1477
Chronic Myelogenous Leukemia with t(9;22)(q34;q11)/BCR-ABL1 Positive 1477
Ph-Negative Myeloproliferative Neoplasms: Polycythemia Vera, Essential Thrombocythemia, and Primary Myelofibrosis—JAK2, MPL Gene, and CALR Mutations 1478
Chronic Neutrophilic Leukemia and Atypical Chronic Myeloid Leukemia with CSF3R Gene Mutation 1480
Mastocytosis and the KIT D816V Gene Mutation 1480
Neoplastic Disorders Associated with Eosinophilia 1480
Molecular Diagnosis of Non-Hodgkin Lymphomas 1481
Rationale for Molecular Genetic Analysis in the Lymphoid Disorders 1481
Antigen Receptor Gene Rearrangements for Clonality Determination 1481
Mechanism of Antigen Receptor Gene Rearrangements 1481
Techniques to Detect Antigen Receptor Gene Rearrangements: Southern Blot Hybridization 1481
Advantages and Shortcomings of Southern Blot Hybridization and Polymerase Chain Reaction for Lymphoid Clonality Assessment 1481
Molecular Detection and Significance of Common Lymphoma-Associated Chromosomal Translocations 1484
t(14;18)/BCL2-IGH@ Abnormality in Follicular and Diffuse Large B Cell Lymphomas 1484
t(11;14)/CCND1-IGH@ Abnormality in Mantle Cell Lymphoma 1485
MALT1, BCL10, and FOXP1 Gene Abnormalities in Extranodal Marginal Zone B Cell Lymphomas 1486
Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma: Molecular and Cytogenetic Prognostic Markers 1486
Diffuse Large B Cell Lymphomas: Role of BCL6 and MYC Gene Abnormalities and Classification Based on Gene Expression Profiling 1487
MYC Gene Translocations in Burkitt Lymphomas and “High-Grade” B Cell Lymphomas, Unclassifiable 1487
MYD88 Mutation in Non-Hodgkin B Cell Lymphoma 1488
ALK Gene Abnormalities in Anaplastic Large Cell Lymphomas 1488
TCL1A Gene Abnormalities in T Cell Prolymphocytic Leukemia 1489
The Implication of Next Gene Sequencing (NGS) in Hematolymphoid Lesions 1489
Emerging Technologies Impacting Molecular Diagnosis and Prognosis Prediction in Hematolymphoid Neoplasia 1489
References 1491
Selected References 1491
References 1491.e1
77 Molecular Genetic Pathology of Solid Tumors 1492
Key Points 1492
Molecular Genetic Pathology of Specific Solid Tumors by Major Organ 1493
Brain Tumor 1493
Glioblastoma Multiforme 1493
Oligodendroglioma 1493
Rhabdoid Tumor 1496
Breast Cancer 1496
Sporadic, Nonhereditary Breast Cancer 1497
Hereditary Breast Cancer 1499
Thyroid Cancer 1499
Papillary Thyroid Carcinoma 1499
Follicular Carcinoma 1500
Medullary Carcinoma 1500
Lung Cancer 1500
Hepatocellular Carcinoma 1501
Hepatic Adenomas 1502
Gastric Cancer 1503
Colorectal Cancer 1504
Hereditary Colonic Cancer Syndrome 1505
Familial Adenomatous Polyposis Syndrome 1505
Hereditary Nonpolyposis Colorectal Cancer (Lynch Syndrome) 1505
Familial Juvenile Polyposis Syndrome 1506
Peutz-Jeghers Syndrome 1506
Serrated Polyposis Syndrome (Formerly Known as Hyperplastic Polyposis Syndrome) 1506
MYH Polyposis and Inherited Colon Cancer Due to MYH Inactivation 1506
Pancreatic Cancer (Non-Neuroendocrine Tumor) 1506
Kidney Tumor 1507
Clear Cell Renal Cell Carcinoma (RCC) 1507
Papillary RCC 1508
Chromophobe Tumors 1508
Micropthalmia Transcription Factor (MiT) Family Translocation Renal Cell Carcinoma 1508
Xp11 Translocation or TFE 3-Associated RCC 1508
(6;11) Translocation TFEB-Associated RCC 1508
Succinate Dehydrogenase B (SDHB) Mutation–Associated RCC 1508
ALK Translocation RCC 1509
Hereditary Renal Cell Carcinoma Syndromes 1509
Gene Expression Profiling by Affymetrix in RCC 1509
Bladder Cancer 1509
Prostate Cancer 1510
Cervical Cancer 1510
Endometrial Cancer 1511
Ovarian Cancer 1511
Histologic Subtypes of Serous Carcinoma 1512
Molecular Classification of Ovarian Cancer 1512
Clear Cell Ovarian Carcinoma 1512
BRCA1 and BRCA2 in Ovarian Cancer 1512
Melanoma 1512
Molecular Genetic Pathology of Sarcoma 1513
Sarcomas with Fusion Genes 1513
Fusion Genes Involving TET Genes 1513
Ewing’s Sarcoma/Primitive Peripheral Neuroectodermal Tumor 1513
Clear Cell Sarcoma 1513
Extraskeletal Myxoid Chondrosarcoma 1513
Atypical Lipomatous Tumor, Well-Differentiated and Dedifferentiated Liposarcoma 1513
Myxoid/Round Cell Liposarcoma 1514
Low-Grade Fibromyxoid Sarcoma 1514
Hemangioma of Bone 1514
Fusion Genes Involving Receptor Tyrosine Kinase Genes 1514
Sarcoma with Fusion Genes Involving Chromatin Remodeling Genes 1514
Sarcoma with Fusion Genes Involving Growth Factors 1514
Sarcoma with Other Types of Fusion Genes 1514
Epithelioid Hemangioendothelioma 1514
Sarcomas with Oncogenic Mutations 1515
KIT and PDGFRA in Gastrointestinal Stromal Tumor 1515
Sarcomas with Variable Other Specific Pattern 1515
Nodular Fasciitis 1515
Inflammatory Myofibroblastic Tumor 1515
Aneurysmal Bone Cyst 1515
Sarcomas with Variable Complex Genetic Alterations with No Specific Pattern 1515
Molecular Genetic Pathology of Multiple Endocrine Neoplasia 1515
Cancers of Unknown Origin 1517
miRNA in Cancer 1518
Clinical Application of Next-Generation Sequencing 1518
References 1519
Selected References 1519
References 1519.e1
78 High-Throughput Genomic and Proteomic Technologies in the Postgenomic Era 1520
Key Points 1520
Overview 1520
The Human Genome Project 1521
Public Sequencing Effort (Hierarchical Shotgun Sequencing) 1521
Private Sequencing Effort (Whole-Genome Shotgun Sequencing) 1522
Finishing the Sequence of the Human Genome 1522
High-Throughput Technologies 1522
Genomic 1522
Serial Analysis of Gene Expression 1523
Microarray 1523
Real Competitive Polymerase Chain Reaction 1524
Multiplex PCR 1527
xTAG Technology 1527
High-Resolution Melting Analysis 1527
Proteomic 1528
Mass Spectrometry 1528
Protein Arrays 1528
Secretome 1528
Molecular Markers for the Diagnosis of Human Neoplasia 1528
Genomic 1528
Proteomic 1529
Diagnostic Cancer Applications 1529
Small, Round Blue Cell Tumors 1529
Barrett’s Esophagus 1530
Thyroid Carcinoma 1530
Distinguishing Follicular Thyroid Carcinoma from Adenoma 1530
Adenocarcinoma 1531
Pancreatic Cancer 1531
Other Cancers 1531
Prognostic Molecular Markers of Disease 1531
Genomic 1531
Diffuse Large B-Cell Lymphoma (see also Chapter 76) 1531
Follicular Lymphoma 1532
Acute Myeloid Leukemia 1532
Breast Cancer 1533
Integrating Genomics and Proteomics 1533
Pitfalls of Molecular Markers for Both Prognostics and Diagnostics 1533
Microarray Data Sets 1533
“Overfitting” The Predictor 1533
The “Bystander” Effect 1534
Limitations of Proteomic Techniques 1534
Conclusion and Future Challenges 1534
References 1534
Selected References 1534
References 1534.e1
Appendix 1 Physiologic Solutions, Buffers, Acid-Base Indicators, Standard Reference Materials, and Temperature Conversions e1
Physiologic Solutions e1
Buffers* e1
Example 1 e1
Example 2 e1
Sorensen’s Phosphate Buffers e1
Tris(hydroxymethyl)aminomethane Buffer* e1
Acid-Base Indicators e1
Standard Reference Materials e2
Temperature Conversion e2
Selected References e4
Appendix 2 Desirable Weights, Body Surface Area, and Body Mass Index e5
Appendix 3 Approximations of Total Blood Volume e9
Adults e9
Children e9
Selected References e9
Appendix 4 Periodic Table of the Elements e10
References e10
Appendix 5 SI Units e11
References e26
Appendix 6 Common Chimeric Genes Identified in Human Malignancies e27
References e29
Appendix 7 Disease/Organ Panels e31
AMA Designated Disease/Organ Panels e32
References e32
Index 1535
A 1535
B 1538
C 1539
D 1543
E 1544
F 1545
G 1546
H 1547
I 1549
J 1551
K 1551
L 1551
M 1553
N 1555
O 1556
P 1557
Q 1559
R 1559
S 1560
T 1562
U 1564
V 1564
W 1565
X 1565
Y 1565
Z 1565
Guidelines for Ordering Blood for Elective Surgery IBC2
Also Referred to as Maximum Surgical Blood Order Schedule (MSBOS) IBC2
Cardiopulmonary/Cardiothoracic IBC2
General Surgery IBC2
Gynecology IBC2
Neurosurgery IBC2
Orthopedics IBC2
Otorhinolaryngology/Head and Neck Surgery IBC3
Plastic Surgery IBC3
Renal Transplantation IBC3
Urology IBC3
Vascular Surgery IBC3
References IBC3