BOOK
Tietz Textbook of Clinical Chemistry and Molecular Diagnostics - E-Book
Carl A. Burtis | Edward R. Ashwood | David E. Bruns
(2012)
Additional Information
Book Details
Abstract
As the definitive reference for clinical chemistry, Tietz Textbook of Clinical Chemistry and Molecular Diagnostics, 5th Edition offers the most current and authoritative guidance on selecting, performing, and evaluating results of new and established laboratory tests. Up-to-date encyclopedic coverage details everything you need to know, including: analytical criteria for the medical usefulness of laboratory procedures; new approaches for establishing reference ranges; variables that affect tests and results; the impact of modern analytical tools on lab management and costs; and applications of statistical methods. In addition to updated content throughout, this two-color edition also features a new chapter on hemostasis and the latest advances in molecular diagnostics.
- Section on Molecular Diagnostics and Genetics contains nine expanded chapters that focus on emerging issues and techniques, written by experts in field, including Y.M. Dennis Lo, Rossa W.K. Chiu, Carl Wittwer, Noriko Kusukawa, Cindy Vnencak-Jones, Thomas Williams, Victor Weedn, Malek Kamoun, Howard Baum, Angela Caliendo, Aaron Bossler, Gwendolyn McMillin, and Kojo S.J. Elenitoba-Johnson.
- Highly-respected author team includes three editors who are well known in the clinical chemistry world.
- Reference values in the appendix give you one location for comparing and evaluating test results.
- NEW! Two-color design throughout highlights important features, illustrations, and content for a quick reference.
- NEW! Chapter on hemostasis provides you with all the information you need to accurately conduct this type of clinical testing.
- NEW! Six associate editors, Ann Gronowski, W. Greg Miller, Michael Oellerich, Francois Rousseau, Mitchell Scott, and Karl Voelkerding, lend even more expertise and insight to the reference.
- NEW! Reorganized chapters ensure that only the most current information is included.
Table of Contents
| Section Title | Page | Action | Price |
|---|---|---|---|
| Front Cover | cover | ||
| Inside Front Cover | ifc1 | ||
| Half title page | i | ||
| Tietz Textbook of Clinical Chemistry and molecular Diagnostics, 5/e | iii | ||
| Copyright Page | iv | ||
| Dedication | v | ||
| Associate Editors | vi | ||
| Reviewers | vi | ||
| Contributors | vii | ||
| Foreword | xv | ||
| Preface | xvi | ||
| Table Of Contents | xvii | ||
| I Principles of Laboratory Medicine | 1 | ||
| 1 Clinical Chemistry, Molecular Diagnostics, and Laboratory Medicine | 3 | ||
| Laboratory Medicine | 3 | ||
| Clinical Chemistry and Laboratory Medicine | 3 | ||
| Ethical Issues in Laboratory Medicine | 4 | ||
| Confidentiality of Genetic Information | 5 | ||
| Confidentiality of Patient Medical Information | 5 | ||
| Allocation of Resources | 5 | ||
| Codes of Conduct | 5 | ||
| Publishing Issues | 5 | ||
| Conflict of Interest | 5 | ||
| The Future | 6 | ||
| References | 6 | ||
| 2 Selection and Analytical Evaluation of Methods—With Statistical Techniques | 7 | ||
| Method Selection | 7 | ||
| Medical Need and Quality Goals | 7 | ||
| Analytical Performance Criteria | 7 | ||
| Other Criteria | 8 | ||
| Basic Statistics | 9 | ||
| Frequency Distribution | 9 | ||
| Population and Sample | 9 | ||
| Probability and Probability Distributions | 9 | ||
| Parameters: Descriptive Measures of a Population | 10 | ||
| Statistics: Descriptive Measures of the Sample | 10 | ||
| Random Sampling | 10 | ||
| The Gaussian Probability Distribution | 10 | ||
| Student t Probability Distribution | 11 | ||
| Nonparametric Statistics | 11 | ||
| Basic Concepts in Relation to Analytical Methods | 12 | ||
| Calibration | 12 | ||
| Trueness and Accuracy | 13 | ||
| Precision | 13 | ||
| Example | 14 | ||
| Precision Profile | 15 | ||
| Linearity | 15 | ||
| Analytical Measurement Range and Limits of Quantification | 16 | ||
| Analytical Sensitivity | 16 | ||
| Analytical Specificity and Interference | 16 | ||
| Analytical Goals | 16 | ||
| Qualitative Methods | 17 | ||
| Performance Measures | 17 | ||
| Agreement Between Qualitative Tests | 18 | ||
| Example | 18 | ||
| Method Comparison | 19 | ||
| Basic Error Model | 19 | ||
| Measured Value, Target Value, Modified Target Value, and True Value | 19 | ||
| Calibration Bias and Random Bias | 20 | ||
| Blunders or Clerical Errors | 21 | ||
| Method Comparison Data Model | 21 | ||
| Comparison of a Routine Method With a Reference Measurement Procedure | 21 | ||
| Comparison of Two Routine Methods | 21 | ||
| Preliminary Practical Work in Relation to a Method Comparison Study | 22 | ||
| Planning a Method Comparison Study | 22 | ||
| Distribution of Differences Plot (DoD plot) | 22 | ||
| Nonparametric Approach | 22 | ||
| Parametric Approach | 24 | ||
| Relative Distribution of Differences Plot (Rel DoD plot) | 24 | ||
| Verification of Distribution of Differences in Relation to Specified Limits | 27 | ||
| Difference (Bland-Altman) Plot | 27 | ||
| A Caution Against Incorrect Interpretation of Paired t-Tests in Method Comparison Studies | 28 | ||
| Regression Analysis | 28 | ||
| Error Models in Regression Analysis | 29 | ||
| Deming Regression Analysis and Ordinary Least-Squares Regression Analysis (OLR) (Constant SDs) | 30 | ||
| Computation Procedures for OLR and Deming Regression | 32 | ||
| Evaluation of the Random Error Around an Estimated Regression Line | 32 | ||
| Interpreting SDy·x (SD21) With Random Errors in Both x1 and x2 | 32 | ||
| Assessment of Outliers | 32 | ||
| The Correlation Coefficient | 33 | ||
| Regression Analysis in Cases of Proportional Random Error | 35 | ||
| Testing for Linearity | 35 | ||
| Nonparametric Regression Analysis (Passing-Bablok) | 35 | ||
| Interpretation of Systematic Differences Between Methods Obtained on the Basis of Regression Analysis | 35 | ||
| Example of Application of Regression Analysis (Weighted Deming Analysis) | 36 | ||
| Discussion of Application of Regression Analysis | 37 | ||
| Monitoring Serial Results | 37 | ||
| Traceability and Measurement Uncertainty | 38 | ||
| Traceability | 38 | ||
| The Uncertainty Concept | 40 | ||
| The Standard Uncertainty (ust) | 40 | ||
| Example of Direct Assessment of Uncertainty on the Basis of Measurements of a Commutable Certified Reference Material | 41 | ||
| Example of Direct Assessment of Uncertainty on the Basis of a Method Comparison Study With a Reference Measurement Procedure Using Patient Samples | 41 | ||
| Indirect Evaluation of Uncertainty by Quantification of Individual Error Source Components | 42 | ||
| Example | 42 | ||
| Uncertainty in Relation to Traditional Systematic and Random Error Classifications | 43 | ||
| Software Packages | 44 | ||
| References | 45 | ||
| 3 Clinical Utility of Laboratory Tests | 49 | ||
| Diagnostic Accuracy of Tests | 49 | ||
| Sensitivity and Specificity | 49 | ||
| Receiver Operating Characteristic Curves | 50 | ||
| Probabilistic Reasoning | 52 | ||
| Prevalence | 52 | ||
| Predictive Values | 52 | ||
| Odds Ratio | 52 | ||
| Likelihood Ratio | 52 | ||
| Bayes’ Theorem | 53 | ||
| Limitations of Bayes’ Theorem | 54 | ||
| Combination Testing | 54 | ||
| Methods For Assessing Diagnostic Accuracy | 56 | ||
| Study Subject Ascertainment | 56 | ||
| Study Design | 56 | ||
| Method of Selection | 57 | ||
| Consecutive/Nonconsecutive Series | 57 | ||
| Test Protocol | 57 | ||
| Verification Procedure | 57 | ||
| Improper Reference Standard | 57 | ||
| Independence | 58 | ||
| Partial Verification | 58 | ||
| Cost-Effectiveness and Outcomes Research | 58 | ||
| References | 58 | ||
| 4 Evidence-Based Laboratory Medicine | 61 | ||
| Concepts, Definitions, and Relationships | 61 | ||
| What Is Laboratory Medicine? | 61 | ||
| What is Evidence-Based Medicine? | 62 | ||
| Definitions, Concepts, and Objectives of Evidence-Based Medicine | 62 | ||
| The Practice of EBM | 63 | ||
| Evidence-Based Medicine and Laboratory Medicine | 64 | ||
| What Is Evidence-Based Laboratory Medicine? | 64 | ||
| The Practice of Evidence-Based Laboratory Medicine | 64 | ||
| Types of Questions Addressed in Laboratory Medicine | 65 | ||
| Formulating an Answerable Question in Laboratory Medicine | 67 | ||
| From Evidence to Outcomes | 68 | ||
| Characterization of the Diagnostic Accuracy of Tests | 68 | ||
| Study Design | 68 | ||
| Reporting of Studies of Diagnostic Accuracy: The Role of the STARD Initiative | 69 | ||
| Using the Test Result | 70 | ||
| Outcome Studies | 70 | ||
| Who Is Interested in Health Outcomes? | 72 | ||
| Test Results Alone Do Not Generate Improved Health Outcomes | 73 | ||
| What Are Outcomes Studies? | 73 | ||
| Design of Clinical Outcomes Studies | 73 | ||
| Comparative Effectiveness | 77 | ||
| Critical Appraisal and Systematic Reviews of Diagnostic Tests | 77 | ||
| Why Systematic Reviews? | 77 | ||
| Conducting a Systematic Review | 78 | ||
| The Clinical Question and Criteria for Selection of Studies | 78 | ||
| Search Strategy | 79 | ||
| Data Extraction and Critical Appraisal of Studies | 79 | ||
| Summarizing the Data | 80 | ||
| Meta-Analysis | 80 | ||
| Economic Evaluation of Diagnostic Tests | 80 | ||
| Who Uses Economic Evaluations of Diagnostic Test? | 80 | ||
| Quality of Evidence in Economic Evaluations | 81 | ||
| Design of Economic Evaluation Studies | 81 | ||
| Choice of Outcome Measures | 82 | ||
| Clinical Outcomes and Economic Evaluation in Decision Making and Changing Practice | 83 | ||
| Clinical Practice Guidelines and Care Pathways | 83 | ||
| What Is a Clinical Guideline? | 83 | ||
| What Is a Care Pathway? | 84 | ||
| The Process of Developing Clinical Guidelines | 84 | ||
| Steps in the Development of Guidelines | 84 | ||
| Selection and Refinement of a Topic | 84 | ||
| Determination of Target Group and Establishment of a Multidisciplinary Guideline Development Team | 84 | ||
| Identifying and Assessing the Evidence | 84 | ||
| Translating Evidence Into a Guideline and Grading the Strength of Recommendations | 84 | ||
| Obtaining External Review and Updating the Guidelines | 86 | ||
| Applying Evidence and Clinical Audit | 87 | ||
| Audit to Help Solve Problems | 87 | ||
| Monitoring Workload and Demand | 88 | ||
| Monitoring the Introduction of a New Test | 88 | ||
| Deleting a Redundant Test | 88 | ||
| Monitoring Adherence to Best Practice | 89 | ||
| Applying the Principles of Evidence-Based Laboratory Medicine in Routine Practice | 89 | ||
| References | 90 | ||
| 5 Establishment and Use of Reference Values | 95 | ||
| The Concept of Reference Values | 95 | ||
| Interpretation by Comparison | 95 | ||
| Normal Values—an Obsolete Term | 95 | ||
| Terminology | 96 | ||
| Clinical Decision Limits | 96 | ||
| Types of Reference Values | 97 | ||
| Subject-Based and Population-Based Reference Values | 97 | ||
| Requirements | 97 | ||
| Selection of Reference Individuals | 97 | ||
| Concept of Health in Relation to Reference Values | 97 | ||
| Strategies for Selection of Reference Individuals | 98 | ||
| Direct or Indirect Sampling? | 98 | ||
| A Priori or A Posteriori Sampling? | 99 | ||
| Random or Nonrandom Sampling? | 100 | ||
| Selection Criteria and Evaluation of Subjects | 100 | ||
| Partitioning of the Reference Group | 101 | ||
| Specimen Collection | 101 | ||
| Preanalytical Standardization | 101 | ||
| Analyte-Specific Considerations | 101 | ||
| The Necessity for Additional Information | 102 | ||
| Analytical Procedures and Quality Control | 103 | ||
| Statistical Treatment of Reference Values | 103 | ||
| Basic Statistical Concepts | 103 | ||
| Sample | 103 | ||
| Estimating Properties | 104 | ||
| Testing Hypotheses | 104 | ||
| Describing the Distribution | 104 | ||
| Reference Limits: Interpercentile Interval | 104 | ||
| Methods Used To Determine Interpercentile Intervals | 105 | ||
| Sample Size | 106 | ||
| Partitioning of Reference Values | 106 | ||
| Inspection of Distribution | 107 | ||
| Identification and Handling of Erroneous Values | 107 | ||
| Methods for Determining Reference Values | 108 | ||
| Nonparametric Method | 108 | ||
| Parametric Method | 108 | ||
| Testing Fit to Gaussian Distribution | 108 | ||
| Transformation of Data: Simple Method | 110 | ||
| Transformation of Data: Two-Stage Method | 110 | ||
| Parametric Estimates of Percentiles and Their Confidence Intervals | 111 | ||
| Other Methods for Calculating Reference Limits | 111 | ||
| Bootstrap Method | 111 | ||
| Robust Method | 111 | ||
| Transferability of Reference Values | 112 | ||
| Analytical Issues | 112 | ||
| Multicenter Trials | 112 | ||
| Verification of Transfer | 112 | ||
| Presentation of An Observed Value in Relation to Reference Values | 112 | ||
| Additional Topics | 113 | ||
| Multivariate, Population-Based Reference Regions | 113 | ||
| The Multivariate Concept | 113 | ||
| The Multiple, Univariate Reference Region | 114 | ||
| The Multivariate Reference Region | 114 | ||
| Subject-Based Reference Values | 115 | ||
| Dynamic versus Static Interpretation of Clinical Chemistry Data | 116 | ||
| References | 116 | ||
| 6 Preanalytical Variables and Biological Variation | 119 | ||
| Preanalytical Variables | 119 | ||
| Controllable Variables | 119 | ||
| Physiologic Variables | 120 | ||
| Posture | 120 | ||
| Prolonged Bed Rest | 121 | ||
| Exercise | 121 | ||
| Physical Training | 122 | ||
| Circadian Variation | 123 | ||
| Travel | 124 | ||
| Diet | 124 | ||
| Food Ingestion | 124 | ||
| Ingestion of Specific Foods and Beverages | 125 | ||
| Bran. | 125 | ||
| Food Constituents. | 125 | ||
| Caffeine. | 125 | ||
| Vegetarianism | 126 | ||
| Malnutrition | 126 | ||
| Long-Term Fasting and Starvation | 127 | ||
| Life-Style | 127 | ||
| Smoking | 127 | ||
| Alcohol Ingestion | 128 | ||
| Drug Administration | 129 | ||
| Herbal Preparations | 130 | ||
| Noncontrollable Variables | 130 | ||
| Biological Influences | 130 | ||
| Age | 131 | ||
| Newborn. | 131 | ||
| Childhood to Puberty. | 132 | ||
| The Adult. | 132 | ||
| The Elderly Adult. | 132 | ||
| Sex | 133 | ||
| Race | 133 | ||
| Environmental Factors | 134 | ||
| Altitude | 134 | ||
| Ambient Temperature | 134 | ||
| Place of Residence | 134 | ||
| Long-Term Cyclical Changes | 134 | ||
| Seasonal Influences | 134 | ||
| Influence of Menstrual Cycle | 135 | ||
| Underlying Medical Conditions | 136 | ||
| Obesity | 136 | ||
| Blindness | 136 | ||
| Pregnancy | 136 | ||
| Stress | 136 | ||
| Fever | 137 | ||
| Shock and Trauma | 137 | ||
| Transfusion and Infusions | 138 | ||
| Biological Variability* | 138 | ||
| Overview | 138 | ||
| Calculating Total Variation | 139 | ||
| Reference Change Values | 140 | ||
| Index of Individuality | 140 | ||
| Index of Heterogeneity | 141 | ||
| Reliability Coefficient | 141 | ||
| Quality Specifications for Total Error Allowable | 141 | ||
| Assessment of Mode of Reporting Test Results | 141 | ||
| References | 142 | ||
| Additional Reading | 144 | ||
| 7 Specimen Collection and Processing | 145 | ||
| Types of Specimens | 145 | ||
| Blood | 145 | ||
| Venipuncture | 145 | ||
| Preliminary Steps | 145 | ||
| Location | 146 | ||
| Preparation of Site | 146 | ||
| Timing | 146 | ||
| Venous Occlusion | 146 | ||
| Order of Draw for Multiple Blood Specimens | 147 | ||
| Collection With Evacuated Blood Tubes | 147 | ||
| Blood Collection With Syringe | 149 | ||
| Completion of Collection | 150 | ||
| Venipuncture in Children | 150 | ||
| Skin Puncture | 150 | ||
| Arterial Puncture | 151 | ||
| Anticoagulants and Preservatives for Blood | 151 | ||
| Heparin | 151 | ||
| Ethylenediaminetetraacetic Acid | 152 | ||
| Sodium Fluoride | 152 | ||
| Citrate | 152 | ||
| Acid Citrate Dextrose | 153 | ||
| Oxalates | 153 | ||
| Iodoacetate | 153 | ||
| Influence of Site of Collection on Blood Composition | 153 | ||
| Collection of Blood from Intravenous or Arterial Lines | 153 | ||
| Hemolysis | 153 | ||
| Urine | 154 | ||
| Timed Urine Specimens | 155 | ||
| Collection of Urine from Children | 155 | ||
| Urine Preservatives | 155 | ||
| Feces | 156 | ||
| Cerebrospinal Fluid | 156 | ||
| Synovial Fluid | 157 | ||
| Amniotic Fluid | 157 | ||
| Chorionic Villus Sampling | 157 | ||
| Pleural, Pericardial, and Ascitic Fluids | 158 | ||
| Saliva | 158 | ||
| Buccal Cells | 158 | ||
| Solid Tissue | 158 | ||
| Hair and Nails | 159 | ||
| Handling of Specimens for Analysis | 159 | ||
| Maintenance of Specimen Identification | 159 | ||
| Preservation of Specimens | 159 | ||
| Separation and Storage of Specimens | 160 | ||
| Transport of Specimens | 160 | ||
| References | 161 | ||
| 8 Quality Management | 163 | ||
| Fundamentals of Total Quality Management | 163 | ||
| Fundamental Concepts | 163 | ||
| Fundamental Principles | 163 | ||
| Total Quality Management of the Clinical Laboratory | 164 | ||
| Establishing Quality Goals and Analytical Performance Limits | 165 | ||
| Laboratory Error and the Six Sigma Process | 166 | ||
| Number of Errors Made in the Clinical Laboratory | 166 | ||
| Six Sigma Principles and Metrics | 167 | ||
| Effect of Analytical Bias on Clinical Decisions | 169 | ||
| Lean Production | 169 | ||
| Elements of a Quality Assurance Program | 169 | ||
| Commitment | 170 | ||
| Facilities and Resources | 170 | ||
| Personnel Competency and Training* | 170 | ||
| Technical Procedures | 171 | ||
| Problem-Solving Mechanism | 172 | ||
| Control of Preanalytical Variables | 172 | ||
| Systems Analysis | 172 | ||
| Types of Preanalytical Variables | 172 | ||
| Test Usage and Practice Guidelines | 173 | ||
| Patient Identification | 173 | ||
| Turnaround Time | 173 | ||
| Transcription Errors | 173 | ||
| Patient Preparation | 173 | ||
| Specimen Collection | 173 | ||
| Specimen Transport | 174 | ||
| Accessioning and Documentation | 174 | ||
| Specimen Separation and Aliquoting | 174 | ||
| Centrifuge Performance | 174 | ||
| Container Monitoring | 174 | ||
| Clerical Errors | 174 | ||
| Control of Analytical Variables | 175 | ||
| Choice of Analytical Method | 175 | ||
| Reference Materials and Methods | 175 | ||
| Definitions of Relevance to Analytical Quality | 175 | ||
| Analytical Traceability | 175 | ||
| Role of International Organization for Standardization (ISO) | 177 | ||
| ISO 9000 | 177 | ||
| ISO Technical Advisory Groups for Laboratory Standards | 177 | ||
| Preparing for ISO Accreditation | 178 | ||
| Documentation of Analytical Protocols | 178 | ||
| Establishment of Reference Intervals | 178 | ||
| Inventory Control of Materials | 178 | ||
| Input Control of Materials | 178 | ||
| Monitoring Method Changes | 178 | ||
| Control of Analytical Quality Using Stable Control Materials and Control Charts | 180 | ||
| Control Materials | 180 | ||
| General Principles of Control Charts | 180 | ||
| Performance Characteristics of a Control Procedure | 182 | ||
| Step-by-Step Process for Selecting QC Procedures | 185 | ||
| Levey-Jennings Control Chart | 185 | ||
| Westgard Multirule Chart | 187 | ||
| Cumulative Sum (Cusum) Control Chart | 188 | ||
| Alternative Calculations and Applications | 191 | ||
| Control of Analytical Quality Using Patient Data | 191 | ||
| Individual Patient Results | 191 | ||
| Clinical Correlation | 191 | ||
| Correlation With Other Laboratory Tests | 192 | ||
| Intralaboratory Duplicates | 192 | ||
| Delta Checks With Previous Test Results | 192 | ||
| Limit Checks | 192 | ||
| Multiple Patients | 193 | ||
| Test Distribution Statistics | 193 | ||
| Statistical Methods for Monitoring Patient Means | 193 | ||
| Combined Use of Liquid Controls and Moving Averages of Patient Values for Quality Control Monitoring | 194 | ||
| External Quality Assessment and Proficiency Testing Programs | 194 | ||
| Quality Control Comparison Programs | 195 | ||
| Proficiency Testing and Laboratory Accreditation | 196 | ||
| Identifying the Sources of Analytical Errors | 197 | ||
| Physical Inspection | 197 | ||
| Relationship of Type and Source of Error | 199 | ||
| Method Evaluation Experiments | 200 | ||
| Optimization Techniques | 200 | ||
| References | 200 | ||
| II Analytical Techniques and Instrumentation | 205 | ||
| 9 Principles of Basic Techniques and Laboratory Safety | 207 | ||
| Concept of Solute and Solvent | 207 | ||
| Expressing Concentrations of Solutions | 207 | ||
| Units of Measurement | 208 | ||
| International System of Units | 208 | ||
| Standardized Reporting of Test Results | 208 | ||
| Lab LOINC System | 209 | ||
| NPU | 209 | ||
| Applications | 209 | ||
| Chemicals | 209 | ||
| Reagent Grade Water | 209 | ||
| Preparation of Reagent Grade Water | 210 | ||
| Distillation | 210 | ||
| Ion Exchange | 210 | ||
| Reverse Osmosis | 210 | ||
| Ultraviolet Oxidation | 210 | ||
| Quality, Use, and Storage of Reagent Grade Water | 210 | ||
| Testing for Water Purity | 210 | ||
| Reagent Grade or Analytical Reagent Grade (AR) Chemicals | 211 | ||
| Ultrapure Reagents | 211 | ||
| Reference Materials | 211 | ||
| Primary Reference Materials | 211 | ||
| Secondary Reference Materials | 211 | ||
| Standard Reference Materials (SRMs) | 211 | ||
| Certified Reference Materials (CRMs) | 211 | ||
| Basic Techniques and Procedures | 213 | ||
| Volumetric Sampling and Dispensing | 213 | ||
| Pipettes | 213 | ||
| Transfer and Measuring Pipettes | 214 | ||
| Transfer Pipettes. | 214 | ||
| Measuring Pipettes. | 214 | ||
| Pipetting Technique | 214 | ||
| Micropipettes | 214 | ||
| Semiautomatic and Automatic Pipettes and Dispensers. | 214 | ||
| Volumetric Flasks | 216 | ||
| Centrifugation | 217 | ||
| Types of Centrifuges | 217 | ||
| Principles of Centrifugation | 217 | ||
| Operation of the Centrifuge | 217 | ||
| Operating Practice | 218 | ||
| Measurement of Radioactivity | 218 | ||
| Basic Concepts | 218 | ||
| Radioactive Decay | 218 | ||
| Alpha Decay. | 218 | ||
| Beta Decay. | 218 | ||
| Electron Capture. | 219 | ||
| Gamma Radiation and Internal Conversion. | 219 | ||
| Activity and Half-life | 219 | ||
| Units of Radioactivity | 219 | ||
| Specific Activity | 220 | ||
| Detection and Measurement of Radioactivity | 220 | ||
| Autoradiography | 220 | ||
| Gas-Filled Detectors | 220 | ||
| Scintillation Counting | 220 | ||
| Crystal Scintillation Detector. | 220 | ||
| Liquid Scintillation Detector. | 220 | ||
| Gravimetry | 220 | ||
| Principles of Weighing | 221 | ||
| Types of Balances | 221 | ||
| Double-Pan Balance | 221 | ||
| Single-Pan Balance | 221 | ||
| Electronic Balance | 221 | ||
| Analytical Weights | 221 | ||
| Thermometry | 221 | ||
| Controlling Hydrogen Ion Concentration | 221 | ||
| Procedures for Processing Solutions | 222 | ||
| Dilution | 222 | ||
| Evaporation | 222 | ||
| Lyophilization | 222 | ||
| Filtration | 222 | ||
| Safety | 223 | ||
| Safety Program | 223 | ||
| Safety Equipment | 223 | ||
| Safety Inspections | 224 | ||
| Plans for the Clinical Laboratory | 224 | ||
| Chemical Hygiene Plan | 224 | ||
| Exposure Control Plan | 224 | ||
| Tuberculosis Control Plan | 225 | ||
| Pandemic Plan | 225 | ||
| Ergonomics Program | 225 | ||
| Hazards in the Laboratory | 225 | ||
| Identification of Hazards | 225 | ||
| Biological Hazards | 226 | ||
| Chemical Hazards | 228 | ||
| Hazards from Volatiles | 228 | ||
| Hazards from Compressed Gases | 229 | ||
| Electrical Hazards | 229 | ||
| Fire Hazards | 229 | ||
| References | 230 | ||
| 10 Optical Techniques | 233 | ||
| Nature of Light | 233 | ||
| Spectrophotometry | 233 | ||
| Basic Concepts | 234 | ||
| Beer’s Law—Relationship Between Transmittance, Absorbance, and Concentration | 234 | ||
| Application of Beer’s Law | 235 | ||
| Instrumentation | 235 | ||
| Components | 236 | ||
| Light Sources | 236 | ||
| Incandescent, Arc, and Cathode Lamps. | 236 | ||
| Laser Sources. | 237 | ||
| Spectral Isolation | 237 | ||
| Filters. | 237 | ||
| Prisms and Gratings. | 238 | ||
| Selection of a Wavelength Isolation Device. | 238 | ||
| Cuvets | 238 | ||
| Photodetectors | 239 | ||
| Photomultiplier Tubes. | 239 | ||
| Photodiodes. | 239 | ||
| Readout Devices | 240 | ||
| Performance Parameters | 240 | ||
| Wavelength Calibration | 240 | ||
| Spectral Bandwidth | 241 | ||
| Stray Light | 241 | ||
| Photometric Accuracy | 241 | ||
| Multiple-Wavelength Readings | 241 | ||
| Reflectance Photometry | 242 | ||
| Flame Emission and Inductively Coupled Plasma Spectrophotometry | 242 | ||
| Atomic Absorption Spectrophotometry | 242 | ||
| Basic Concepts | 242 | ||
| Instrumentation | 242 | ||
| Interferences in Atomic Absorption Spectrophotometry | 243 | ||
| Spectral Interferences | 243 | ||
| Nonspectral Interferences | 244 | ||
| Fluorometry | 244 | ||
| Basic Concepts | 244 | ||
| Time Relationships of Fluorescence Emission | 244 | ||
| Relationship of Concentration and Fluorescence Intensity | 245 | ||
| Fluorescence Polarization | 245 | ||
| Instrumentation | 246 | ||
| Components | 246 | ||
| Excitation Source | 246 | ||
| Xenon Lamp. | 246 | ||
| Lasers. | 246 | ||
| Excitation and Emission Monochromator | 247 | ||
| Cuvet | 247 | ||
| Photodetectors | 248 | ||
| Visual Observation. | 248 | ||
| Photomultiplier Tube. | 248 | ||
| Charge-Coupled Detector. | 248 | ||
| Performance Verification | 248 | ||
| Types of Fluorometers and Spectrofluorometers | 249 | ||
| Ratio-Referencing Spectrofluorometer | 249 | ||
| Time-Resolved Fluorometer | 250 | ||
| Flow Cytometer | 250 | ||
| Hematofluorometer | 251 | ||
| Limitations of Fluorescence Measurements | 251 | ||
| Inner Filter Effects | 251 | ||
| Concentration Quenching | 251 | ||
| Light Scattering | 251 | ||
| Cuvet Material and Solvent Effects | 252 | ||
| Sample Matrix Effects | 252 | ||
| Temperature Effects | 252 | ||
| Photodecomposition | 252 | ||
| Phosphorescence | 252 | ||
| Chemiluminescence, Bioluminescence, and Electrochemiluminescence | 252 | ||
| Basic Concepts | 252 | ||
| Chemiluminescence and Bioluminescence | 253 | ||
| Electrochemiluminescence | 253 | ||
| Instrumentation | 253 | ||
| Limitations of Chemiluminescence and Electrochemiluminescence Measurements | 253 | ||
| Nephelometry and Turbidimetry | 253 | ||
| Basic Concepts | 253 | ||
| Particle Size | 253 | ||
| Wavelength Dependence of Light Scattering | 254 | ||
| Concentration and Molecular Weight Factors in Light Scattering | 254 | ||
| Effects of Polarized Light on Light Scattering | 254 | ||
| Angular Dependence of Light Scattering | 255 | ||
| Light Scattering and Plasma Proteins | 255 | ||
| Measurement of Scattered Light | 255 | ||
| Turbidimetry | 255 | ||
| Nephelometry | 255 | ||
| Selection of Method | 255 | ||
| Instrumentation | 255 | ||
| Turbidimeter | 255 | ||
| Nephelometer | 256 | ||
| Limitations of Light Scattering Measurements | 256 | ||
| Antigen Excess | 256 | ||
| Matrix Effects | 257 | ||
| References | 257 | ||
| 11 Electrochemistry and Chemical Sensors | 259 | ||
| Potentiometry and Ion-Selective Electrodes | 259 | ||
| Basic Concepts | 259 | ||
| Types of Electrodes | 260 | ||
| Redox Electrodes | 260 | ||
| Inert Metal Electrodes | 260 | ||
| Metal Electrodes Participating in Redox Reactions | 261 | ||
| Ion-Selective Electrodes | 261 | ||
| The Glass Electrode | 262 | ||
| Polymer Membrane Electrodes | 262 | ||
| Electrodes for PCO2 | 265 | ||
| Direct Potentiometry by ISE—Units of Measure and Reporting for Clinical Applications | 266 | ||
| Voltammetry/Amperometry | 267 | ||
| Basic Concepts | 267 | ||
| Applications | 269 | ||
| Conductometry | 271 | ||
| Coulometry | 272 | ||
| Optical Chemical Sensors | 272 | ||
| Basic Concepts | 273 | ||
| Applications | 273 | ||
| Biosensors | 274 | ||
| Enzyme-Based Biosensors With Amperometric Detection | 274 | ||
| Enzyme-Based Biosensors With Potentiometric and Conductometric Detection | 277 | ||
| Enzyme-Based Biosensors With Optical Detection | 277 | ||
| Affinity Sensors | 278 | ||
| Chemical Sensors Based On Nanotechnology | 281 | ||
| In Vivo and Minimally Invasive Sensors | 281 | ||
| References | 283 | ||
| 12 Electrophoresis | 287 | ||
| Basic Concepts And Definitions | 287 | ||
| Theory Of Electrophoresis | 287 | ||
| Conventional Electrophoresis | 289 | ||
| Instrumentation | 289 | ||
| Power Supplies | 289 | ||
| Buffers | 289 | ||
| Support Media | 290 | ||
| Starch Gel | 290 | ||
| Cellulose Acetate | 290 | ||
| Agarose | 290 | ||
| Polyacrylamide Gel | 291 | ||
| Automated Systems | 291 | ||
| General Operations | 291 | ||
| Electrophoretic Separation | 291 | ||
| Detection and Quantification | 291 | ||
| Staining | 291 | ||
| Quantification | 292 | ||
| Blotting Techniques | 292 | ||
| Technical and Practical Considerations | 293 | ||
| Sampling | 293 | ||
| Discontinuities in Sample Application | 293 | ||
| Unequal Migration Rates | 293 | ||
| Distorted, Unusual, or Atypical Bands | 293 | ||
| Types of Electrophoresis | 293 | ||
| Slab Gel Electrophoresis | 293 | ||
| Disc Electrophoresis | 294 | ||
| Isoelectric Focusing Electrophoresis | 294 | ||
| Isotachophoresis | 295 | ||
| Pulsed-Field Electrophoresis | 295 | ||
| Two-Dimensional (2D) Electrophoresis | 295 | ||
| Capillary Electrophoresis | 296 | ||
| Instrumentation | 296 | ||
| The Capillary Format | 296 | ||
| Sample Injection | 297 | ||
| Direct Detection | 297 | ||
| Improving Limits of Detection | 297 | ||
| Increased OPL. | 297 | ||
| Sensitive Optical Detectors. | 297 | ||
| Online Sample Concentration. | 297 | ||
| Indirect Detection | 298 | ||
| Technical Considerations | 298 | ||
| Temperature Effects | 298 | ||
| Surface Effects | 298 | ||
| Modes of Operation | 299 | ||
| Capillary Zone Electrophoresis | 299 | ||
| Serum Protein Analysis | 299 | ||
| Artifacts in Serum Protein Analysis | 299 | ||
| Micellar Electrokinetic Chromatography (MEKC) | 300 | ||
| Capillary Gel Electrophoresis | 300 | ||
| Capillary Isoelectric Focusing Electrophoresis | 300 | ||
| Capillary Isotachophoresis | 301 | ||
| Microchip Electrophoresis | 301 | ||
| Instrumentation | 301 | ||
| Fabrication of Microchips | 301 | ||
| Molecular Diagnostics Using Microchips | 302 | ||
| References | 303 | ||
| 13 Chromatography and Extraction | 307 | ||
| Chomatography | 307 | ||
| Separation Mechanisms | 307 | ||
| Ion-Exchange | 308 | ||
| Adsorption | 309 | ||
| Partition Chromatography | 310 | ||
| Chiral Separations | 311 | ||
| Size Exclusion Chromatography | 311 | ||
| Other Size Exclusion Separations | 312 | ||
| Affinity Chromatography | 312 | ||
| Resolution | 312 | ||
| Retention Factor | 313 | ||
| Efficiency (N) | 314 | ||
| Selectivity | 314 | ||
| Peak Capacity | 315 | ||
| Planar Chromatography | 315 | ||
| Liquid Chromatography | 315 | ||
| Instrumentation | 315 | ||
| Solvent Reservoir | 315 | ||
| Solvent Delivery System | 315 | ||
| Sample Injector | 316 | ||
| Column Heaters/Chillers | 317 | ||
| Columns | 317 | ||
| Column Dimensions. | 317 | ||
| Column Stationary Phase. | 317 | ||
| Particulate Column Packings. | 317 | ||
| Bonded Phase Packings. | 318 | ||
| Polymeric Packings. | 318 | ||
| Chiral Packings. | 318 | ||
| Restricted Access Packings. | 318 | ||
| Monolithic Columns. | 318 | ||
| Detectors | 318 | ||
| Photometers and Spectrophotometers. | 318 | ||
| Fluorometers. | 319 | ||
| Electrochemical Detectors. | 320 | ||
| Refractive Index Detectors. | 320 | ||
| Mass Spectrometry. | 320 | ||
| Other Detectors. | 320 | ||
| System Controller and Data System | 320 | ||
| Safety | 321 | ||
| Supercritical Fluid Chromatography (SFC) | 321 | ||
| Gas Chromatography | 321 | ||
| Instrumentation | 321 | ||
| Columns | 321 | ||
| Carrier Gas Supply and Flow Control | 322 | ||
| Injector | 322 | ||
| Temperature Control | 323 | ||
| Detectors | 323 | ||
| Flame Ionization Detector. | 323 | ||
| Thermionic Selective Detector. | 324 | ||
| Electron Capture Detector. | 324 | ||
| Photoionization Detector. | 325 | ||
| Thermal Conductivity Detector. | 325 | ||
| Mass Spectrometry. | 325 | ||
| Computer/Controller | 325 | ||
| Qualitative and Quantitative Analyses | 325 | ||
| Analyte Identification | 325 | ||
| Analyte Quantification | 326 | ||
| Extraction and Differential Precipitation | 326 | ||
| References | 327 | ||
| 14 Mass Spectrometry | 329 | ||
| Basic Concepts and Definitions | 329 | ||
| Instrumentation | 331 | ||
| Ion Source | 331 | ||
| Electron Ionization | 332 | ||
| Chemical Ionization | 332 | ||
| Electrospray Ionization | 332 | ||
| Atmospheric Pressure Chemical Ionization | 333 | ||
| Atmospheric Pressure Photoionization | 334 | ||
| Inductively Coupled Plasma | 334 | ||
| Matrix-Assisted Laser Desorption/Ionization | 334 | ||
| Ionization Methods of Potential Interest | 334 | ||
| Ionization Methods of Historical Interest | 335 | ||
| Vacuum System | 335 | ||
| Mass Analyzers, Tandem Mass Spectrometers, and Ion Detectors | 335 | ||
| General Classes of Mass Spectrometers | 335 | ||
| Beam-Type Designs | 335 | ||
| Quadrupole. | 336 | ||
| Magnetic Sectors. | 337 | ||
| Time-of-Flight. | 337 | ||
| Trapping Mass Spectrometers | 338 | ||
| Quadrupole Ion Trap. | 338 | ||
| Linear Ion Trap. | 339 | ||
| Ion Cyclotron Resonance. | 339 | ||
| Orbitrap. | 340 | ||
| Tandem Mass Spectrometers | 340 | ||
| Detectors | 341 | ||
| Computer and Software | 342 | ||
| Clinical Applications | 342 | ||
| Gas Chromatography–Mass Spectrometry | 343 | ||
| Liquid Chromatography–Mass Spectrometry | 343 | ||
| MALDI MASS Spectrometry | 345 | ||
| ICP Mass Spectrometry | 346 | ||
| Proteomics, Genomics, and Metabolomics | 346 | ||
| Practical Aspects of Mass Spectrometry—Logistics, Operations, and Quality | 348 | ||
| References | 350 | ||
| 15 Enzyme and Rate Analyses | 355 | ||
| Basic Principles | 355 | ||
| Enzyme Nomenclature | 355 | ||
| Enzymes as Proteins | 355 | ||
| Specificity and the Active Center | 357 | ||
| Isoenzymes and Other Multiple Forms of Enzymes | 357 | ||
| Genetic Origins of Enzyme Variants | 358 | ||
| Nongenetic Causes of Multiple Forms of Enzymes | 358 | ||
| Distribution of Isoenzymes and Other Multiple Forms of Enzymes | 360 | ||
| Changes in Isoenzyme Distribution During Development and Disease | 360 | ||
| Differences in Properties Between Multiple Forms of Enzymes | 361 | ||
| Enzymes as Catalysts | 361 | ||
| Units for Expressing Enzyme Activity | 361 | ||
| Enzyme Kinetics | 362 | ||
| The Enzyme–Substrate Complex | 362 | ||
| Factors Governing the Rate of Enzyme-Catalyzed Reactions | 362 | ||
| Enzyme Concentration | 362 | ||
| Substrate Concentration | 363 | ||
| Single-Substrate Reactions | 363 | ||
| Two-Substrate Reactions | 365 | ||
| Consecutive Enzymatic Reactions | 366 | ||
| Effect of pH | 366 | ||
| Temperature | 367 | ||
| Inhibitors and Activators | 368 | ||
| Inhibition of Enzyme Activity | 368 | ||
| Reversible Inhibition. | 368 | ||
| Irreversible Inhibition. | 369 | ||
| Inhibition by Antibodies. | 370 | ||
| Enzyme Activation | 370 | ||
| Coenzymes and Prosthetic Groups | 370 | ||
| Analytical Enzymology | 371 | ||
| Measurement of Reaction Rates | 371 | ||
| Measurement of Substrates | 372 | ||
| Optimization, Standardization, and Quality Control | 373 | ||
| Optimization | 373 | ||
| Standardization | 373 | ||
| Quality Control | 374 | ||
| Measurement of Enzyme Mass Concentration | 374 | ||
| Enzymes as Analytical Reagents | 374 | ||
| Measurement of Metabolites | 374 | ||
| Equilibrium Methods | 374 | ||
| Kinetic Methods | 375 | ||
| Immunoassay | 375 | ||
| Analytical Applications of Immobilized Enzymes | 375 | ||
| Measurement of Isoenzymes and Isoforms | 375 | ||
| Electrophoresis | 375 | ||
| Chromatography | 376 | ||
| Chemical Inactivation and Differences in Catalytic Properties | 376 | ||
| Immunochemical Assays | 376 | ||
| References | 376 | ||
| 16 Principles of Immunochemical Techniques | 379 | ||
| Basic Concepts | 379 | ||
| Antibodies | 379 | ||
| Immunogens | 379 | ||
| Antigen-Antibody Binding | 381 | ||
| Binding Forces | 381 | ||
| Van der Waals-London Dipole-Dipole Interactions | 381 | ||
| Hydrophobic Interaction | 381 | ||
| Coulombic Bonds | 381 | ||
| Reaction Mechanism | 381 | ||
| Factors Influencing Binding | 382 | ||
| Ion Species and Ionic Strength Effects | 382 | ||
| Polymer Effect | 382 | ||
| Types of Reactions | 382 | ||
| The Precipitin Reaction | 382 | ||
| Reactions at a Solid-Liquid Interface | 383 | ||
| Qualitative Methods | 384 | ||
| Passive Gel Diffusion | 384 | ||
| Immunoelectrophoresis (IEP) | 384 | ||
| Crossed Immunoelectrophoresis (CRIE) | 384 | ||
| Counterimmunoelectrophoresis (CIE) | 385 | ||
| Immunofixation (IF) | 385 | ||
| Western Blotting | 385 | ||
| Dot Blotting | 386 | ||
| Quantitative Methods | 386 | ||
| Radial Immunodiffusion and Electroimmunoassay | 386 | ||
| Radial Immunodiffusion Immunoassay | 386 | ||
| Electroimmunoassay | 387 | ||
| Turbidimetric and Nephelometric Assays | 387 | ||
| Labeled Immunochemical Assays | 388 | ||
| Methodologic Principles | 388 | ||
| Competitive versus Noncompetitive Reaction Formats | 388 | ||
| Competitive Immunoassays. | 388 | ||
| Noncompetitive Immunoassays. | 389 | ||
| Heterogeneous versus Homogeneous Immunochemical Assays | 390 | ||
| Heterogeneous Assays. | 390 | ||
| Homogeneous Assays. | 390 | ||
| Analytical Detection Limits | 390 | ||
| Examples of Labeled Immunoassays | 391 | ||
| Radioimmunoassay | 391 | ||
| Enzyme Immunoassay (EIA) | 391 | ||
| Enzyme-Linked Immunosorbent Assay. | 392 | ||
| Enzyme Multiplied Immunoassay Technique. | 392 | ||
| Cloned Enzyme Donor Immunoassay. | 393 | ||
| Fluoroimmunoassay | 393 | ||
| Chemiluminescence Immunoassay | 394 | ||
| Electrochemiluminescence Immunoassay | 395 | ||
| Simplified Immunoassays | 395 | ||
| Protein Microarrays | 396 | ||
| Interferences in Immunoassays | 396 | ||
| Cell and Tissue-Based Immunochemical Techniques | 397 | ||
| Immunohistochemistry | 397 | ||
| Agglutination Assays | 397 | ||
| References | 397 | ||
| 17 Nucleic Acid Techniques | 401 | ||
| Enzymes That Act On Nucleic Acids | 401 | ||
| Nucleic Acid Treatments that Do Not Use Enzymes | 402 | ||
| Amplification Techniques | 403 | ||
| Polymerase Chain Reaction (PCR)—Target Amplification | 403 | ||
| Details of the PCR Process | 403 | ||
| PCR Kinetics and Rapid Cycling | 405 | ||
| PCR Optimization and Primer Design | 405 | ||
| Detection Limits of PCR | 406 | ||
| 3′-End of PCR Products | 406 | ||
| Contamination Control to Avoid False-Positive Results | 407 | ||
| Inhibition Control to Avoid False-Negative Results | 407 | ||
| Hot Start Techniques | 408 | ||
| Asymmetric PCR and Allele-Specific PCR | 408 | ||
| Single-Molecule PCR or Digital PCR | 408 | ||
| Other Forms of Target Amplification | 408 | ||
| Transcription-Based Amplification Methods | 408 | ||
| Strand Displacement Amplification | 409 | ||
| Loop-Mediated Amplification Methods | 409 | ||
| Whole Genome and Whole Transcriptome Amplification | 409 | ||
| Other Approaches to Amplification | 409 | ||
| Branched-Chain DNA: Signal Amplification | 409 | ||
| Serial Invasive Amplification: Signal Amplification | 410 | ||
| Rolling Circle Amplification: Probe Amplification | 410 | ||
| End Point Quantification in Amplification Assays | 410 | ||
| Detection Techniques | 410 | ||
| Generic Measurement and Visualization of Nucleic Acids | 410 | ||
| UV Absorbance | 410 | ||
| Fluorescent Staining of Nucleic Acids | 410 | ||
| Reporter Molecules and Labeled Probes | 411 | ||
| Radioactivity | 411 | ||
| Indirect Probe Detection | 411 | ||
| Fluorescent Labels | 411 | ||
| Discrimination Techniques | 412 | ||
| Electrophoresis | 412 | ||
| Restriction Fragment Length Polymorphism | 413 | ||
| Southern and Northern Blotting | 413 | ||
| PCR Product Length | 414 | ||
| PCR/RFLP | 414 | ||
| Heteroduplex Migration | 414 | ||
| Single-Strand Conformation Polymorphism | 415 | ||
| Denaturing Gradient Gel Electrophoresis | 416 | ||
| Temperature Gradient Electrophoresis | 416 | ||
| Dideoxy-Termination Sequencing | 416 | ||
| Single Nucleotide Extension (SNE) | 418 | ||
| Oligo Ligation | 418 | ||
| Multiplex Ligation-Dependent Probe Amplification | 419 | ||
| Alternatives to Electrophoresis | 419 | ||
| Pyrosequencing | 419 | ||
| Mass Spectrometry | 419 | ||
| High-Performance Liquid Chromatography (HPLC) | 420 | ||
| High-Throughput Sequencing | 420 | ||
| Sequencing by Ligation | 421 | ||
| High-Throughput Sequencing Without Amplification | 422 | ||
| Hybridization Assays: Principles | 422 | ||
| Hybridization Thermodynamics | 422 | ||
| Hybridization Kinetics | 424 | ||
| Probes | 425 | ||
| Cloned Probes | 425 | ||
| PCR-Generated Probes | 425 | ||
| Oligonucleotide Probes | 425 | ||
| Estimating Tms of Oligonucleotide Probes | 425 | ||
| Purity of Labeled Oligonucleotide Probes | 425 | ||
| Hybridization Assays: Examples | 426 | ||
| Dot-Blot and Line-Probe Assays | 426 | ||
| Medium-Density Arrays | 426 | ||
| Microarrays | 427 | ||
| Single-Nucleotide Polymorphism (SNP) Arrays | 427 | ||
| Gene Expression Arrays | 427 | ||
| Copy Number Variant Arrays | 428 | ||
| In Situ Hybridization | 428 | ||
| Single-Copy Visualization | 429 | ||
| Real-Time PCR | 429 | ||
| Dyes and Probe Formats for Real-Time PCR | 431 | ||
| Double-Stranded DNA Binding Dyes | 431 | ||
| Fluorescently Labeled Primers | 431 | ||
| Probe-Specific Detection | 432 | ||
| Hybridization Probes. | 432 | ||
| Hydrolysis Probes. | 432 | ||
| Mixed Mechanism Probes. | 434 | ||
| Detection and Quantification in Real-Time PCR | 434 | ||
| Detection | 434 | ||
| Quantification | 435 | ||
| Accuracy and Precision | 436 | ||
| Melting Analysis | 436 | ||
| SNP Genotyping | 437 | ||
| High-Resolution Melting Analysis | 437 | ||
| Comparison of Closed-Tube SNP Genotyping Methods | 437 | ||
| Conclusion | 439 | ||
| References | 439 | ||
| 18 Microfabrication and Microfluidics and Their Application to Clinical Diagnostics | 443 | ||
| Background | 443 | ||
| Microdevice Fabrication | 443 | ||
| Photolithography and Glass Etching Techniques | 444 | ||
| Alternatives to Traditional Photolithography and Wet Chemical Etching | 445 | ||
| Polymeric Materials | 445 | ||
| Polydimethylsiloxane (PDMS) | 445 | ||
| Plexiglas (PMMA) | 446 | ||
| Separation and Detection of Clinically Relevant Analytes | 447 | ||
| Electrophoretic Separations | 447 | ||
| Detection on Microchips | 448 | ||
| Exemplary Separations | 449 | ||
| Separation of Nucleic Acids | 449 | ||
| Mutation Detection via Microchip Analysis | 450 | ||
| Other Mutation Detection Schemes | 452 | ||
| Separation of Proteins | 453 | ||
| Separation of Small Molecules and Ions | 454 | ||
| Microfluidic Valving | 456 | ||
| Applications | 456 | ||
| Nucleic Acids | 456 | ||
| DNA Extraction | 456 | ||
| Polymerase Chain Reaction | 457 | ||
| Integration | 457 | ||
| Hybridization Arrays | 457 | ||
| Proteins | 458 | ||
| Immunoassays | 458 | ||
| Protein Microarrays | 460 | ||
| Cells | 460 | ||
| Detection of Drugs and Drug Metabolites | 462 | ||
| Limitations of Microfluidic Systems | 462 | ||
| Future of Microfluidics in Clinical Diagnostics | 463 | ||
| References | 463 | ||
| 19 Automation in the Clinical Laboratory | 469 | ||
| Basic Concepts | 469 | ||
| Automation of the Analytical Processes | 470 | ||
| Specimen Identification | 470 | ||
| Labeling | 470 | ||
| Bar Coding | 471 | ||
| Identification Errors | 471 | ||
| Specimen Preparation | 471 | ||
| Use of Whole Blood for Analysis | 471 | ||
| Automation of Specimen Preparation | 472 | ||
| Specimen Delivery | 472 | ||
| Pneumatic Tube Systems | 472 | ||
| Electric Track Vehicles | 472 | ||
| Mobile Robots | 472 | ||
| Specimen Loading and Aspiration | 472 | ||
| Sample Pretreatment | 473 | ||
| Sample Introduction and Internal Transport | 473 | ||
| Discrete Processing Systems | 473 | ||
| Carryover | 473 | ||
| Reagent Handling and Storage | 474 | ||
| Reagent Identification | 474 | ||
| Open versus Closed Systems | 474 | ||
| Reagent Delivery | 474 | ||
| Chemical Reaction Phase | 474 | ||
| Type of Reaction Vessel and Cuvet | 475 | ||
| Mixing of Reactants | 475 | ||
| Thermal Regulation | 475 | ||
| Separation in Immunoassay Systems | 475 | ||
| Measurement Approaches | 475 | ||
| Photometry/Spectrophotometry | 475 | ||
| Optical Source | 475 | ||
| Spectral Isolation | 475 | ||
| Photometric Detectors | 475 | ||
| Reflectance Photometry | 476 | ||
| Fluorometry | 476 | ||
| Turbidimetry and Nephelometry | 476 | ||
| Chemiluminescence and Bioluminescence | 476 | ||
| Electrochemical | 477 | ||
| Signal Processing, Data Handling, and Process Control | 477 | ||
| Test Autoverification | 477 | ||
| Integrated Automation for the Clinical Laboratory | 478 | ||
| Workstations | 478 | ||
| Instrument Clusters | 478 | ||
| Work Cells | 478 | ||
| Automated Specimen Processing | 479 | ||
| Stand-Alone Specimen Processing Systems | 479 | ||
| Integrated and Modular Automation Systems | 479 | ||
| Conveyor Belts | 480 | ||
| Automated Specimen Sorting | 482 | ||
| Automated Specimen Storage and Retrieval | 482 | ||
| Practical Considerations | 482 | ||
| Evaluation of Requirements | 482 | ||
| Problems of Integration | 483 | ||
| Device Integration | 483 | ||
| Other Areas of Automation | 483 | ||
| Urine Analyzers | 483 | ||
| Cell Counters | 484 | ||
| Nucleic Acid Analyzers | 484 | ||
| Microtiter Plate Systems | 484 | ||
| Automated Pipetting Stations | 484 | ||
| References | 484 | ||
| 20 Point-of-Care Testing | 487 | ||
| Analytical and Technological Solutions | 487 | ||
| Hand-Held, Single-Use Devices | 488 | ||
| Urine and Blood Dipsticks | 489 | ||
| Glucose Strips and Meters | 489 | ||
| Coagulation Strips and Meters | 491 | ||
| Lateral Flow Strips | 492 | ||
| Cartridge or Cassette Devices | 493 | ||
| Multiple-Use Cartridges and Benchtop Systems | 493 | ||
| Informatics and Poct | 495 | ||
| Informatic Requirements in POCT Devices | 495 | ||
| Development of POCT Connectivity Standards | 496 | ||
| Benefits of Connectivity and Future Developments | 496 | ||
| Implementation and Management Considerations | 497 | ||
| Establishment of Need, Risks, And Change Management Challenges | 497 | ||
| Organization and Implementation of a Coordinating Committee | 498 | ||
| Poct Policy and Accountability | 498 | ||
| Equipment Procurement and Evaluation | 498 | ||
| Training and Certification Of Operators | 499 | ||
| Quality Control, Quality Assurance, and Audit | 500 | ||
| Maintenance and Inventory Control | 501 | ||
| Documentation | 501 | ||
| Accreditation and Regulation of Poct | 502 | ||
| Future of Poct | 502 | ||
| References | 503 | ||
| III Analytes | 507 | ||
| 21 Amino Acids, Peptides, and Proteins | 509 | ||
| Amino Acids | 510 | ||
| Basic Biochemistry | 510 | ||
| Acid-Base Properties of Amino Acids | 510 | ||
| Hydrophobicity, Solubility, and Stability of Amino Acids | 510 | ||
| Amino Acid Metabolism | 513 | ||
| Amino Acid Concentrations | 514 | ||
| Clinical Implications of Amino Acid Concentrations | 515 | ||
| Analysis of Amino Acids | 516 | ||
| Peptides and Proteins | 516 | ||
| Peptide and Protein Structure | 516 | ||
| Disorders of Protein Folding | 517 | ||
| Protein Synthesis and Processing | 518 | ||
| Physical Properties of Proteins | 518 | ||
| Plasma Proteins | 519 | ||
| The Plasma Proteome and Peptidome | 519 | ||
| Plasma Protein Concentrations | 521 | ||
| Major Plasma Proteins | 521 | ||
| The Acute-Phase Response | 523 | ||
| Differences Between Plasma and Serum Proteins | 524 | ||
| Determination of Total Protein | 525 | ||
| Specific Methods. | 525 | ||
| Kjeldahl Method. | 525 | ||
| Biuret Method. | 525 | ||
| Direct Optical Methods. | 525 | ||
| Dye-Binding Methods. | 525 | ||
| Lowry (Folin-Ciocalteu) Method. | 526 | ||
| Refractometry. | 526 | ||
| Turbidimetric and Nephelometric Methods. | 526 | ||
| Calibration of Total Protein Methods. | 526 | ||
| Reference Intervals. | 526 | ||
| Prealbumin (Transthyretin) and Retinol-Binding Protein (RBP) | 526 | ||
| Biochemistry and Function. | 526 | ||
| Clinical Significance of Prealbumin and RBP. | 527 | ||
| Increased Plasma Concentrations. | 527 | ||
| Decreased Plasma Concentrations. | 527 | ||
| Genetic Aspects. | 527 | ||
| Laboratory Considerations and Reference Intervals. | 527 | ||
| Albumin | 527 | ||
| Biochemistry of Albumin. | 527 | ||
| Function of Albumin. | 528 | ||
| Clinical Significance of Albumin | 528 | ||
| Increased Plasma Concentrations. | 528 | ||
| Decreased Plasma Concentrations. | 528 | ||
| Analbuminemia. | 528 | ||
| Inflammation. | 528 | ||
| Hepatic Disease. | 528 | ||
| Urinary Loss/Kidney Disease. | 528 | ||
| Gastrointestinal Loss. | 528 | ||
| Protein-Calorie Malnutrition. | 528 | ||
| Burn Injury. | 529 | ||
| Edema and Ascites. | 529 | ||
| Genetic Aspects of Albumin. | 529 | ||
| Laboratory Considerations for Albumin | 529 | ||
| Plasma and Serum. | 529 | ||
| Reference Intervals of Albumin. | 529 | ||
| α1-Acid Glycoprotein (AAG) | 530 | ||
| Biochemistry of AAG. | 530 | ||
| Function of AAG. | 530 | ||
| Clinical Significance of AAG. | 530 | ||
| Increased Plasma Concentrations | 530 | ||
| Acute-Phase Response. | 530 | ||
| IV Molecular Diagnostics and Genetics | 1207 | ||
| 37 Principles of Molecular Biology | 1209 | ||
| Landmark Developments in Genetics and Molecular Diagnostics | 1209 | ||
| The Essentials | 1210 | ||
| Nucleic Acid Structure and Organization | 1210 | ||
| Molecular Compositions and Structures of DNA and RNA | 1210 | ||
| DNA | 1210 | ||
| RNA | 1211 | ||
| Chromosome Structure | 1211 | ||
| Chromatin Packing | 1212 | ||
| Nucleic Acid Physiology and Functional Regulation | 1214 | ||
| Replication | 1214 | ||
| Transcription | 1215 | ||
| Translation | 1216 | ||
| Genetics and Epigenetics | 1216 | ||
| DNA Methylation | 1217 | ||
| Histone Modifications | 1218 | ||
| Nucleosome Positioning and Remodeling | 1218 | ||
| Noncoding RNAs | 1219 | ||
| MicroRNA | 1219 | ||
| Other ncRNAs | 1219 | ||
| Beyond the Nuclear Genome | 1220 | ||
| The Mitochondrial Genome | 1220 | ||
| Circulating Nucleic Acids | 1220 | ||
| Understanding Our Genome | 1220 | ||
| References | 1222 | ||
| 38 Genomes and Nucleic Acid Alterations | 1225 | ||
| Human Genome | 1225 | ||
| Variation Within the Human Genome | 1226 | ||
| Variations That Cause Human Disease | 1227 | ||
| Single-Nucleotide Polymorphisms | 1227 | ||
| Small Insertions and/or Deletions | 1227 | ||
| Structural Variants | 1227 | ||
| Haplotypes | 1227 | ||
| Alterations in Hemizygous Genes and Mitochondrial DNA | 1227 | ||
| Human Epigenetic Alterations | 1228 | ||
| Bacterial Genomes | 1228 | ||
| Viral Genomes | 1228 | ||
| Fungal Genomes | 1229 | ||
| References | 1229 | ||
| Genome Databases | 1229 | ||
| Human Genes and Disease | 1229 | ||
| Sequence Databanks | 1229 | ||
| Human Variation Databases | 1229 | ||
| Nomenclature | 1229 | ||
| 39 Nucleic Acid Isolation | 1231 | ||
| Specimen Preservation | 1231 | ||
| Tissue Homogenization and Cell Lysis | 1232 | ||
| DNA Isolation | 1232 | ||
| RNA Isolation | 1232 | ||
| Assessment of Nucleic Acid Yield and Quality | 1233 | ||
| Storage of Purified Nucleic Acids | 1233 | ||
| Automated Nucleic Acid Isolation | 1233 | ||
| Point-of-Care Nucleic Acid Analysis | 1235 | ||
| Isolation of Circulating Nucleic Acids | 1235 | ||
| References | 1235 | ||
| 40 Inherited Diseases | 1239 | ||
| Diseases with Mendelian Inheritance | 1239 | ||
| Autosomal Recessive Diseases | 1239 | ||
| Cystic Fibrosis | 1240 | ||
| Hereditary Hemochromatosis | 1243 | ||
| Spinal Muscular Atrophy | 1245 | ||
| Autosomal Dominant Diseases | 1247 | ||
| Achondroplasia | 1247 | ||
| Huntington’s Disease | 1248 | ||
| Marfan Syndrome | 1252 | ||
| X-Linked Diseases | 1253 | ||
| Hemophilia A | 1253 | ||
| Duchenne Muscular Dystrophy | 1255 | ||
| Fragile X Syndrome | 1257 | ||
| Diseases with Nonmendelian Inheritance | 1259 | ||
| Mitochondrial DNA Diseases | 1259 | ||
| Leber Hereditary Optic Neuropathy | 1261 | ||
| Leigh Syndrome | 1261 | ||
| Imprinting | 1262 | ||
| Prader-Willi and Angelman Syndromes | 1262 | ||
| Complex Diseases | 1264 | ||
| Thrombophilia | 1265 | ||
| Inherited Breast Cancer | 1266 | ||
| Inherited Colon Cancer | 1269 | ||
| Reporting of Test Results | 1273 | ||
| Glossary | 1274 | ||
| References | 1275 | ||
| 41 Identity Assessment | 1293 | ||
| Variation in the Human Genome | 1293 | ||
| Genetic Variation Useful in Identity Testing | 1293 | ||
| Exclusion of Tested Individuals | 1294 | ||
| Likelihood of Inclusion of Tested Individuals | 1295 | ||
| Samples Employed for Identity Testing | 1295 | ||
| Forensic Dna Typing | 1295 | ||
| Forensic Applications | 1295 | ||
| Genetic Systems Used in Forensic Identification | 1296 | ||
| VNTR Analysis by RFLP | 1296 | ||
| Short Tandem Repeats | 1296 | ||
| Gender Markers and Y Chromosome Markers | 1297 | ||
| Mitochondrial DNA | 1297 | ||
| Single-Nucleotide Polymorphisms | 1298 | ||
| Other Systems | 1298 | ||
| Instrumentation Used in Forensic Laboratories | 1298 | ||
| Quality Assurance and Accreditation in Forensic DNA Analysis | 1298 | ||
| Statistical Interpretation | 1299 | ||
| Convicted Offender Databases | 1299 | ||
| Legal Issues | 1299 | ||
| Use of Dna Testing for the Identification of Clinical Specimens | 1300 | ||
| Transplantation Testing | 1300 | ||
| Genetic Features of HLA Genes | 1300 | ||
| Historical Applications of HLA Typing | 1301 | ||
| Forensic DNA Testing | 1301 | ||
| Parentage Testing | 1302 | ||
| Chimerism | 1302 | ||
| Transplantation | 1302 | ||
| DNA-Based HLA Allele Identification | 1302 | ||
| Interpretation of HLA Test Results | 1303 | ||
| Quality Assurance and Quality Control Issues | 1304 | ||
| Proficiency Testing | 1304 | ||
| Accreditation and Certification of Laboratories and Professionals | 1304 | ||
| Chimerism and Hematopoietic Cell Engraftment Analysis | 1304 | ||
| Methods for Performing Engraftment Analysis | 1304 | ||
| Selection and Interpretation of Short Tandem Repeat Loci | 1305 | ||
| Applications of Microsatellite Locus Testing to Engraftment Analysis | 1305 | ||
| Proficiency Testing | 1306 | ||
| Quality Assurance and Quality Control | 1306 | ||
| Accreditation | 1306 | ||
| Parentage Testing | 1306 | ||
| Methods, Instrumentation, and Sample Requirements | 1306 | ||
| Reporting of Test Results | 1306 | ||
| Test Reports | 1306 | ||
| Exclusion of a Tested Man | 1306 | ||
| Inclusion of a Tested Man | 1307 | ||
| Opinions, Interpretations, and Problems | 1307 | ||
| Quality Assurance and Quality Control | 1307 | ||
| Policy and Procedures for Resolution of Complaints | 1307 | ||
| Accreditation | 1307 | ||
| References | 1308 | ||
| World Wide Web Sites | 1308 | ||
| 42 Molecular Methods in Diagnosis and Monitoring of Infectious Diseases | 1309 | ||
| Chlamydia Trachomatis and Neisseria Gonorrhoeae | 1310 | ||
| Nucleic Acid Testing for CT and GC | 1310 | ||
| False-Positive Test Results | 1311 | ||
| False-Negative Test Results | 1312 | ||
| Liquid Cytology Specimens | 1312 | ||
| Recommendations on Laboratory Testing for CT and GC | 1312 | ||
| Human Papillomavirus | 1312 | ||
| Nucleic Acid Testing for HPV | 1313 | ||
| Human Immunodeficiency Virus Type 1 | 1314 | ||
| HIV-1 Viral Load Testing | 1314 | ||
| Available HIV-1 Viral Load Assays | 1315 | ||
| Qualitative and Proviral HIV-1 RNA Testing | 1316 | ||
| HIV-1 Resistance Testing | 1316 | ||
| HIV-1 Tropism Testing | 1318 | ||
| Herpes Simplex Virus | 1318 | ||
| Nucleic Acid Testing for HSV | 1318 | ||
| Enterovirus 2 | 1319 | ||
| Nucleic Acid Testing for Enteroviruses | 1319 | ||
| Perinatal Group B Streptococcal Disease | 1320 | ||
| Nucleic Acid Testing for GBS | 1320 | ||
| Cytomegalovirus | 1321 | ||
| Nucleic Acid Testing for CMV | 1321 | ||
| Mycobacterium Tuberculosis | 1323 | ||
| Nucleic Acid Testing for MTb | 1323 | ||
| Hepatitis C Virus | 1324 | ||
| Detection and Quantification of HCV | 1324 | ||
| Available Assays for Detection and Quantification | 1325 | ||
| HCV Genotyping | 1326 | ||
| Clostridium Difficile | 1326 | ||
| Nucleic Acid Tests for C. difficile | 1327 | ||
| Methicillin-Resistant Staphylococcus Aureus (Mrsa) | 1327 | ||
| Nucleic Acid Testing for MRSA | 1328 | ||
| Respiratory Viruses | 1329 | ||
| Nucleic Acid Testing for Respiratory Viruses | 1330 | ||
| References | 1332 | ||
| 43 Pharmacogenetics | 1339 | ||
| Defining Pharmacogenetic Targets | 1339 | ||
| Approaches to Pharmacogenetic Testing | 1341 | ||
| Clinical Application of Pharmacogenetic Testing | 1344 | ||
| Pharmacogenetics in Oncology | 1345 | ||
| Pharmacogenetics in Psychiatry and Neurology | 1345 | ||
| Pharmacogenetics in Cardiology | 1347 | ||
| Pharmacogenetics in Infectious Disease | 1347 | ||
| Phase I Metabolic Enzymes: Cytochrome P450 Isozymes | 1348 | ||
| Cytochrome P450 2D6 (CYP2D6) | 1348 | ||
| Genotype to Phenotype | 1348 | ||
| Testing | 1351 | ||
| Tamoxifen Application | 1351 | ||
| Codeine Application | 1353 | ||
| Antidepressant Application | 1353 | ||
| Cytochrome P450 2C9 (CYP2C9) | 1354 | ||
| Genotype to Phenotype | 1354 | ||
| Testing | 1354 | ||
| Warfarin Application | 1355 | ||
| Cytochrome P450 2C19 (CYP2C19) | 1355 | ||
| Genotype to Phenotype | 1355 | ||
| Testing | 1356 | ||
| Clopidogrel Application | 1356 | ||
| Antidepressant Application | 1356 | ||
| Omeprazole and Proguanil Applications | 1356 | ||
| Other Cytochrome P450 Genes | 1357 | ||
| Phase II Metabolic Enzymes | 1357 | ||
| N-Acetyltransferases (NAT1 and NAT2) | 1357 | ||
| Genotype to Phenotype | 1357 | ||
| Testing | 1358 | ||
| Clinical Applications | 1359 | ||
| Thiopurine S-Methyltransferase | 1359 | ||
| Genotype to Phenotype | 1359 | ||
| Testing | 1360 | ||
| 6-Mercaptopurine and Azathioprine Applications | 1361 | ||
| UDP-Glucuronosyltransferase 1A1 (UGT1A1) | 1361 | ||
| Genotype to Phenotype | 1362 | ||
| Irinotecan Application | 1362 | ||
| Protease Inhibitors Application | 1362 | ||
| Other Phase II Metabolism Enzymes | 1363 | ||
| Pharmacodynamic Genes | 1363 | ||
| Vitamin K Epoxide Reductase Complex 1 (VKORC1) | 1363 | ||
| Genotype to Phenotype | 1363 | ||
| Warfarin Sensitivity Application | 1363 | ||
| Human Leukocyte Antigen (HLA) Complex, Class I, B | 1364 | ||
| Genotype to Phenotype | 1364 | ||
| Risk of SCARS and Related Drug Hypersensitivity Applications | 1364 | ||
| Future Directions | 1365 | ||
| References | 1365 | ||
| 44 Hematopoietic Malignancies | 1371 | ||
| Antigen Receptor Rearrangements for Determination of Clonality | 1371 | ||
| Molecular Genetic Basis for Immunoglobulin Gene Rearrangements | 1371 | ||
| Molecular Genetic Basis for T-Cell Receptor Gene Rearrangements | 1374 | ||
| Southern Blot Hybridization Analysis for Antigen Receptor Gene Rearrangements | 1374 | ||
| Polymerase Chain Reaction Analysis of Antigen Receptor Gene Rearrangements | 1376 | ||
| Molecular Genetics of Malignant Lymphomas | 1377 | ||
| Southern Blot Hybridization Analysis for the Detection of Chromosomal Translocations | 1378 | ||
| Polymerase Chain Reaction Analysis for Detection of Chromosomal Translocations | 1378 | ||
| t(14;18)(q32;21)—BCL2/JH Aberration | 1378 | ||
| t(11;14)(q13;32)—CCND1/JH Aberration* | 1379 | ||
| Molecular Genetics of Leukemias | 1380 | ||
| Recurrent Chromosomal Translocations in Acute Myeloid Leukemias | 1380 | ||
| t(8;21)(q22;q22)—AML1-ETO | 1382 | ||
| t(15;17)(q22;q21)—PRAM1 (also known as PML-RARA) | 1383 | ||
| Recurrent Chromosomal Translocations in Chronic Leukemias of Myeloid/Monocytic Lineage | 1383 | ||
| t(9;22)(q34;q11)—BCR-ABL Aberration | 1383 | ||
| t(9:22) in ALL | 1386 | ||
| Reverse Transcription Polymerase Chain Reaction | 1386 | ||
| Real-Time Polymerase Chain Reaction | 1386 | ||
| Quantification | 1387 | ||
| ABL-1 Drug Resistance Mutations in Ph+ Chromosome–Positive Leukemias | 1387 | ||
| Mutations of Oncogenes and Tumor Suppressor Genes in Hematopoietic Malignancies | 1387 | ||
| Malignant Lymphomas | 1387 | ||
| A20/TNFAIP3 | 1388 | ||
| Acute Myeloid Leukemias | 1388 | ||
| RAS Mutations | 1388 | ||
| FMS-like Tyrosine Kinase 3 | 1388 | ||
| Nucleophosmin 1 (NPM1) Gene Mutations | 1388 | ||
| CCAAT Enhancer–Binding Protein Alpha (Cebpα) Gene Mutations | 1389 | ||
| Partial Tandem Duplications (PTDs) of the Myeloid/Lymphoid or Mixed Lineage Leukemia (MLL) Gene | 1389 | ||
| Wilms’ Tumor Suppressor 1 Gene (WT1) | 1389 | ||
| Myeloproliferative Disorders | 1389 | ||
| Janus Kinase (JAK) 2 Gene Mutations | 1389 | ||
| Myeloproliferative Leukemia Virus Oncogene Homology (MPL) Gene Mutations | 1390 | ||
| Minimal Residual Disease Detection and Monitoring | 1390 | ||
| Detection of Viral Genomes | 1391 | ||
| Epstein-Barr Virus | 1391 | ||
| In Situ Hybridization | 1392 | ||
| Conclusions | 1392 | ||
| References | 1392 | ||
| 45 Plasma Nucleic Acids | 1397 | ||
| Discovery and Early Work | 1397 | ||
| Circulating DNA as a Tumor Marker | 1397 | ||
| Measurement of Total Plasma DNA Concentrations | 1397 | ||
| Plasma DNA Size as a Tumor Marker | 1397 | ||
| Detection of Loss of Heterozygosity in Plasma | 1398 | ||
| Detection of Somatic Gene Mutations in Plasma | 1398 | ||
| Detection of Aberrant DNA Methylation in Plasma | 1399 | ||
| Viral DNA in Plasma | 1399 | ||
| Circulating RNA as a Tumor Marker | 1400 | ||
| Fetal Nucleic Acids in Maternal Plasma | 1400 | ||
| Discovery and Basic Biology of Fetal DNA in Maternal Plasma | 1400 | ||
| Applications to Sex-Linked Diseases, RhD Genotyping, and Monogenic Disease | 1401 | ||
| Detection of Chromosomal Aneuploidies | 1402 | ||
| Pregnancy-Associated Disorders | 1404 | ||
| Other Applications of Plasma Nucleic Acids | 1405 | ||
| Concluding Remarks | 1405 | ||
| References | 1405 | ||
| V Pathophysiology | 1413 | ||
| 46 Diabetes Mellitus | 1415 | ||
| Classification | 1415 | ||
| Type 1 Diabetes Mellitus | 1416 | ||
| Type 2 Diabetes Mellitus | 1416 | ||
| Other Specific Types of Diabetes Mellitus | 1416 | ||
| Gestational Diabetes Mellitus | 1416 | ||
| Impaired Glucose Tolerance | 1416 | ||
| Impaired Fasting Glucose | 1416 | ||
| Hormones that Regulate Blood Glucose Concentration | 1416 | ||
| Insulin | 1417 | ||
| Chemistry | 1417 | ||
| Synthesis | 1417 | ||
| Release | 1419 | ||
| Degradation | 1419 | ||
| Proinsulin | 1420 | ||
| C-Peptide | 1420 | ||
| Antibodies to Insulin | 1420 | ||
| The Mechanism of Insulin Action | 1420 | ||
| Glucose Transport | 1422 | ||
| Insulin-like Growth Factors | 1422 | ||
| Counter-Regulatory Hormones | 1423 | ||
| Glucagon | 1423 | ||
| Epinephrine | 1423 | ||
| Growth Hormone | 1423 | ||
| Cortisol | 1423 | ||
| Other Hormones Influencing Glucose Metabolism | 1423 | ||
| Thyroxine | 1423 | ||
| Somatostatin | 1423 | ||
| Clinical Utility of Measuring Insulin, Proinsulin, C-Peptide, and Glucagon | 1424 | ||
| Insulin | 1424 | ||
| Proinsulin | 1424 | ||
| C-Peptide | 1425 | ||
| Fasting Hypoglycemia | 1425 | ||
| Insulin Secretion | 1425 | ||
| Monitoring Therapy | 1425 | ||
| Glucagon | 1425 | ||
| Methods for the Measurement of Specific Hormones | 1425 | ||
| Insulin | 1425 | ||
| Principle | 1426 | ||
| Comments | 1426 | ||
| VI Appendix | 2129 | ||
| 60 Reference Information for the Clinical Laboratory | 2131 | ||
| Contents | 2131 | ||
| Index | 2189 | ||
| A | 2189 | ||
| B | 2193 | ||
| C | 2195 | ||
| D | 2200 | ||
| E | 2203 | ||
| F | 2204 | ||
| G | 2206 | ||
| H | 2208 | ||
| I | 2212 | ||
| J | 2214 | ||
| K | 2214 | ||
| L | 2215 | ||
| M | 2217 | ||
| N | 2220 | ||
| O | 2221 | ||
| P | 2222 | ||
| Q | 2228 | ||
| R | 2228 | ||
| S | 2229 | ||
| T | 2232 | ||
| U | 2235 | ||
| V | 2236 | ||
| W | 2237 | ||
| X | 2238 | ||
| Y | 2238 | ||
| Z | 2238 | ||
| Inside Back Cover | ibc1 |