Manual of Molecular and Clinical Laboratory Immunology: 2 Volume Set (ASM Books)

THE authoritative guide for clinical laboratory immunology For nearly 50 years, the Manual of Molecular and Clinical Laboratory Immunology has been the premier resource for laboratories, students, and professionals involved in the clinical and technical details of diagnostic immunology testing. The 9th Edition continues its tradition of providing comprehensive clinical and technical information on the latest technologies used in medical and diagnostic immunology. Led by a world-renowned group of authors and editors, this new edition reflects substantial changes aimed at improving and updating the Manual’s utility while reflecting the significant transformations that have occurred since the last edition, including the revolution of gene editing and the widespread adoption of molecularly engineered cellular therapies. Topical highlights include: Laboratory Management: three new chapters cover essential aspects of quality assurance, quality improvement, and quality management, aligning with the increasingly stringent and demanding regulatory environment. Inborn Errors of Immunity: the primary immunodeficiency section has been completely updated to align with the latest International Union of Immunological Societies’ classifications of inborn errors of immunity. Functional Cellular Assays: expanded content includes detailed discussions on various functional assays critical for modern immunologic testing. Autoimmune Diseases: expanded chapters on systemic and organ-specific autoimmune disorders, including new chapters on Sjögren’s syndrome and deficiency of ADA2, as well as significant updates on organ-specific autoimmune diseases. Transplantation Immunology: updated chapters detail the assessment of immune reconstitution and ABO testing, reflecting latest practices. The 9th Edition of the Manual of Molecular and Clinical Laboratory Immunology serves as an invaluable resource for laboratory directors, clinicians, laboratory managers, technologists, and students. It provides critical insights into the selection, application, and interpretation of immunologic tests, offering practical guidance on troubleshooting, clinical application, and an understanding of test limitations. This comprehensive and up-to-date manual remains an essential tool for anyone involved in the diagnosis, evaluation, and management of immune-mediated and immune system-related disorders. Cover Volume 1 Title Page Copyright Page Contents Editorial Board Contributors Preface Author and Editor Conflicts of interest Section A Molecular Methods Chapter 1 Introduction References Chapter 2 Molecular Methods for Genetic Diagnosis of Inborn Errors of Immunity Introduction Sequencing Technique Progression Sanger Sequencing High-Throughput Sequencing Pre- and postsequencing Considerations Sample Selection Analysis of variants Additional Techniques Copy Number Variation Trecs Droplet Digital PCR Future Considerations Acknowledgments References Chapter 3 Immunoglobulin and T-cell Receptor Gene Assessment Introduction Protein Structure of immunoglobulin Basic Immunoglobulin Structure Heavy-chain Isotypes Structure of the B-cell Receptor Protein Structure of the T-cell Receptor (tcr) T- and B-cell Development T-cell Development B Cell Development Genetic Structure of Immunoglobulins and t-cell Receptor Immunoglobulin Genes T-cell Receptor Genes Immunoglobulin and t-cell Receptor Gene Rearrangement B- and T-cell Receptor Gene Rearrangement Mechanism CDR3 Diversity J (joining) Region C (constant) Region Recombination Signal Sequences IGHC Class Switching Somatic Hypermutation Productive/unproductive Rearrangement Molecular Methods for TCR and IG Rearrangement Testing PCR-based Assays NGS-based Assays Considerations In assay Design Quality Controls for NGS Clonality Assays Interpretation of TCR and IG Rearrangement Testing Clonality Diagnostics and MRD Testing MRD Diagnostics Using Cell-free DNA Conclusions References Section B Immunoglobulin Methods Chapter 4 Introduction References Chapter 5 Clinical Indications and applications of Serum and Urine Protein Electrophoresis Background on protein Structure Principles of Protein Electrophoresis Zone Electrophoresis Quality Control and quality Assurance Specimen Requirements for Serum Specimen Requirements for Urine Internal Controls Proteins Identified In Serum Protein Electrophoresis Pattern Interpretation In Serum Liver Disease Pattern Protein Loss Patterns Acute-phase Reaction Detection of M-proteins In Serum Quantification of Serum M-proteins Pattern Interpretation In Urine Detection of M-Proteins In the Urine Clinical Applications Avoiding False-positive Results Correlation of current With previous Patient Results References Chapter 6 Immunochemical Characterization of immunoglobulins In Serum, Urine, and cerebrospinal Fluid Background Laboratory Investigation Immunofixation Immunosubtraction Pattern Interpretations and clinical Disorders Polyclonal Endogenous Monoclonal Proteins Monoclonal Gammopathies Diseases With monoclonal Immunoglobulin Deposition Heavy Chain Diseases Plasma Cell Neoplasms Exogenous (therapeutic) Monoclonal Proteins and chimeric Proteins Oligoclonal Banding Hypogammaglobulinemia Artifacts Quality Control Urine Immunofixation Cerebrospinal Fluid Laboratory Investigation References Chapter 7 Cryoglobulins and Cryofibrinogenemia Cryoglobulins Background Concept Procedures Isolation, Quantitation, and characterization of cryoglobulins Assays for specific Antibody Activities, Antigens, or Detection of nucleic Acid Complement Measurements In cryoglobulinemia Conclusions Intralaboratory Quality Control and interlaboratory Proficiency Testing Cryofibrinogens Background Concept Interpretation Procedures Conclusions Anticoagulant-related Recommendations References Chapter 8 Strategy for detecting and following Monoclonal Gammopathies Classification of monoclonal Gammopathies Malignant Plasma Cell Proliferative Disorders Premalignant Plasma Cell Proliferative Diseases Protein (or Low-tumor-burden) Diseases Overview of laboratory Tests for monoclonal Gammopathies Diagnostic Testing Strategies for M-Proteins In symptomatic Patients Monitoring M-proteins Stratification of risk Summary References Section C Complement Chapter 9 Introduction References Chapter 10 Methods of complement Analysis Introduction Preanalytical Considerations Diagnostic Workup Methods of modern Complement Analysis Functional Assays Protein Quantification of individual Components Analysis of complement Activation Products Multiplex Analysis of complement Proteins Monitoring of complement-targeted Drugs Complement Analysis In tissues Quality Assessment Outlook References Chapter 11 Assessing Complement Dysregulation In the Thrombotic Microangiopathies and ultrarare Complement-mediated Kidney Disease Alternative Pathway of complement Complement-mediated Renal Disease Atypical Hemolytic Uremic Syndrome C3 Glomerulopathy Conclusions Acknowledgements References Chapter 12 Current and future of complement Therapeutics and their Impact On the Clinical Immunology Laboratory C1-Inihibitor Concentrates Anti-C5 Antibodies Eculizumab and ABP959 Ravulizumab Crovalimab Pozelimab Anti-C5a Antibody Anti-masp-2 Antibody Cyclic Peptide C3 inhibitor Small-molecule C5a Receptor Antagonist Anti-C1 Antibodies Oral Small-molecule Inhibitors Pegylated RNA Aptamers RNA Interference Adeno-associated Virus (AAV) Gene therapy References Section D Flow Cytometry Chapter 13 Introduction: Clinical Flow Cytometry Applications and quality Control References Chapter 14 Polychromatic Flow Cytometry Technology Instrumentation Overview Lasers Optical Filters Signal Processing and configuration of instrument Detectors Flow Cytometer Standardization Sample Preparation Sample Types and requirements Reagents Procedures Panel Design, One Approach Doublet Exclusion Quality Assurance and quality Control Requirements for validation of an Assay Include: Data Analysis and interpretation Future Directions References Chapter 15 High-sensitivity Detection of Paroxysmal Nocturnal Hemoglobinuria by Multiparameter Flow Cytometry Introduction Clinical Features of pnh Who Should Be tested for pnh? Complement Blockade Therapy and disease Monitoring In pnh Early Methods To test for pnh Evolution of flow Cytometric Methods To detect Pnh Issues With early Flow Methods Routine Versus High-sensitivity Flow Cytometry High-sensitivity Assays for pnh Red and white Blood Cells General Requirements High-sensitivity 2-c Assay for pnh In mature Rbcs Antibody Clone/conjugate Selection for High-sensitivity Detection of pnh MRBCs Instrument Setup Considerations for MRBCs MRBC Staining Protocol Using CD235a FITC/CD59 Pe Cocktail Issues With MRBC Assays Assays for pnh Within Immature Rbcs Cross-platform 3-color (3-c) Assay for pnh Irbcs and MRBCs Validation of 3-color Irbc/MRBC Assay To screen Pnh Samples High-sensitivity 4-Color (4-C) WBC assays Antibody Clone/conjugate Selection for 4-C High-sensitivity Pnh WBC Detection Instrument Setup Considerations: WBCs WBC Staining Procedure for optimized Neutrophil and monocyte Cocktails High-sensitivity 5-Color (5-C) WBC assays Antibody Clone/conjugate Selection for 5-C High-sensitivity Pnh WBC Detection Instrument Setup Considerations for 5-Color WBC assay Staining Procedure for 5-Color High-sensitivity Combined Neutrophil/monocyte Assay Presence of type Ii Populations In neutrophils and monocytes CD157-negative Cases High-sensitivity 6-Color (6-C) WBC assays Antibody Clone/conjugate Selection for 6-C High-sensitivity Pnh WBC Detection IS FLAER Essential for high-sensitivity Testing of pnh WBCs? Cross-platform 5-, 6-, and 7-color Single Tube Assays for pnh WBCs Considerations for verification of high-sensitivity Pnh Assays Verification of instrument Setup and antibody Performance Internal Controls To verify Instrument Setup and antibody Conjugate Performance Fluorescence-minus-two (fm2) Controls To verify Instrument Settings Routine Control of assay Quality Considerations for validation of high-sensitivity Pnh Assays Ongoing QC and qc Monitors Reporting References Chapter 16 Standardized Flow Cytometry Assays for Enumerating CD34+ Hematopoietic Stem Cells Introduction Hematopoietic Stem/progenitor Cells Express CD34 Antigen Transplantation of hematopoietic Stem Cells Measuring Graft Adequacy Graft Assessment by CD34+ Cell Enumeration and Flow Cytometry Clinical Issues In enumerating Stem Cells by Flow Cytometry Early Methods to Measure CD34+ Cells by Flow Cytometry Statistical Issues In rare Event Detection Appropriate Controls for rare Event Detection Technical Issues in CD34+ Cell Enumeration by Multiparameter Flow Cytometry CD34 Antigen: Structural Considerations and choice of fluorochromes CD45 Antigen: Structural Considerations and choice of fluorochromes CD34+ Cell Enumeration Using Sequential Boolean Gating The Basic Ishage Protocol and gating Strategy Single-platform Absolute CD34+ Cell Counting Ishage Single-platform Including Viability Assessment Ishage-Based Commercial Kits for CD34+ Cell Enumeration Benefits of Single-platform CD34+ Cell enumeration Negative Antibody Controls Lysing Agents Issues In the Analysis of postthawed Samples Quality Assurance of CD34+ Cell enumeration Simultaneous Enumeration of CD34+ and CD3+ Cells in the Allogeneic Transplant Setting Immunological Characterization of CD34+ Stem Cells Does Enumerating CD34+ Subsets Have Clinical Utility? CD34+ Cell Subsets in Backup Marrow of Poor Mobilizers Use of plerixafor In patients Who Fail To mobilize With g-csf References Chapter 17 the Detection of fetal Red Cells In fetomaternal Hemorrhage by Flow Cytometry Introduction Application of the Assay Procedure Sample Requirement Materials/reagents/commercial Controls Safety Equipment/instrumentation Procedure Sample Setup and antibody Staining Frbc Panel Acquisition On the Flow Cytometer Postacquisition Analysis Calculations Quality Control/assurance Fetaltrol Level 1 (negative), Level 2 (low Positive), and level 3 (high Positive) Whole-blood Controls Negative-normal Whole-blood Specimen (preferably Male or Nonpregnant Female) Interpretation Expected Values Analytical Sensitivity Analytical Specificity Acknowledgments References Chapter 18 Functional Flow Cytometry-based Assays of myeloid and lymphoid Functions for the Diagnostic Screening of inborn Errors of immunity Flow Cytometric Evaluation of phosphorylated Kinase Substrates To evaluate Abnormalities In immune Signaling Pathways Induction and measurement of stat1 Phosphorylation Levels As a Screen for type 1 Cytokine Signaling Abnormalities Associated With primary Immunodeficiency Flow Cytometric Determination of stat1 gain-of-function Alleles In patients With cmCD Cell Surface Adhesion Marker Upregulation As a Screen for lad-1 Lad-1, CD18 Deficiency Flow Cytometric Assessment of cell Surface Adhesion Marker Upregulation As a Screen for lad-1 Measurement of the Upregulation of CD40 ligand On activated T Cells As a Screening Diagnostic Test for CD40 ligand Deficiency (formally Known As x-linked Hyper IGm Syndrome [xhim]) Measurement of CD40 ligand Expression on in Vitro-activated T-helper Cells and CD40 Receptor Expression on B Cells, Respectively, as Screening Diagnostic Tests for CD40 ligand Deficiency and CD40 Receptor Deficiency A Flow Cytometry-based Oxidative Burst Assay As a Screen for the Diagnosis of cgd Chronic Granulomatous Disease The Oxidative Burst Assay As a Screening Test for the Diagnosis of cgd Measurement of abnormal T Cell and nk Cell Cytotoxicity As an Aid in the Diagnosis of patients Suspected of hemophagocytic Lymphohistiocytosis Syndromes Measurement of cell Surface CD107a As a Surrogate of the Degranulation Process Involved In t Cell and nk Cell Cytotoxicity Measurement of cell Surface CD278 Expression for the Diagnosis of icos Deficiency Summary References Chapter 19 Quality Assurance and validation In clinical Flow cytometry Introduction Regulation, Accredidation, and quality Management Standardization and guidelines Procedural Code Standardization Requirements Prior To assay Development and validation Instrument Qualification Installation Qualification (iq) Operational Qualification (oq) Performance Qualification (pq) Reagent Qualification Staff Qualification Verification Validation Validation Report Quality Assurance (qa) Quality Control (qc) Procedures Reagent QC Method/assay QC Patient Internal QC Specimen Integrity/quality Assessment Cell Quantity and optimal Adjustment Document Control System (DCS) External Quality Assessment (EQA) Quality Audits Staff Training and competency Occurrence Management Quality Improvement (QI) Conclusion References Chapter 20 Future Cytometric Technologies and applications Introduction Pros and cons of the Technologies Number of parameters Spillover/spectral Overlap Sensitivity Acquisition Speed Sorting Cost Controls Best Applications for each Technology Imaging Cytometry As another New frontier References Section E Functional Cellular Assays Chapter 21 Introduction Chapter 22 Cryopreservation of blood Mononuclear Cells Technical Aspects of pbmc Cryopreservation, Storage, and transportation Cryopreservation and thawing Protocol Cell Quality Required for immunologic Assays of cryopreserved Pbmcs Lymphoproliferative Assays T Cell Elispot Assays B Cell Elispot Assays Functional and phenotypic Assays Using Flow Cytometry Molecular Assays On cryopreserved Pbmcs Quality Control and advances In the Processing and storage of cryopreserved Pbmcs CAVD/EQAPOL Long-term Pbmc Repository: Maintenance of t Cell Functionality As measured by Elispot Assay Effects of cell Concentration of cryopreserved Preparations and hiv Status On viable Recovery Effect of leukopak Processing On functional Response References Chapter 23 Functional Assays for b Cells and antibodies Approaches for Assessing B Cell Function In Vivo B Cell Function In Vitro B-cell Function Methodology: in Vitro Whole-blood Lymphocyte Proliferation Assay Approaches for assessing Antibody Function Bactericidal Assays Methodology: Serum Bactericidal Assay for Functional Antibodies Against Haemophilus Influenzae Type B Opsonophagocytosis Assays (opas) Methodology: Multiplexed Opsonophagocytic Killing Assay (mopa4) for Functional Antibodies Against Streptococcus Pneumoniae Acknowledgments References Chapter 24 Methods for detection of antigen-specific T Cells by Enzyme-linked Immunospot Assay Introduction Enzyme-linked Immunospot Assay (elispot) Protocol Day 1: Coating Elispot Plates and Thawing Cryopreserved Pbmc Samples Day 2: Addition of cells and stimulating Antigen Day 3: Assay Development Methods of enhancing Elispot Sensitivity Positive Controls Interpreting Elispot Data: Implementing Quality Controls and Identifying Positive Responses Research and Clinical Applications of the Elispot assay References Chapter 25 Regulatory T Cell (treg) Characterization: Phenotype, Function, and clinical Importance of human Treg Human CD4+ Regulatory T Cells Characterisation of Human CD4+ Treg Methods To identify and quantify Ttreg Methods To identify and quantify Tr1 Cells Methods to Test the Function of CD4+ Treg Treg and disease References Chapter 26 Measurement of natural Killer Cell Phenotype and activity in humans Introduction Procedures Sample Requirements Materials and reagents Equipment and instrumentation Experimental Procedure Controls Quality Control and assurance Pitfalls and troubleshooting Interpretation Conclusions References Chapter 27 Functional Assays for the Diagnosis of chronic Granulomatous Disease Introduction Analysis of Pmn H2o2 Production by Flow Cytometry of Dihydrorhodamine 123 Staining Principle Isolation and characterization of pmn Principle Histochemical Staining of pmn With nitroblue Tetrazolium Principle Quantitative Analysis of Generation Using Superoxide Dismutase-inhibitable Ferricytochrome C Reduction Principle Analysis of pmn Ros Generation By luminol-enhanced Chemiluminescence Principle Analysis of Gp91phox Surface Expression by Flow Cytometry Principle Flow Cytometric Analysis of phox Protein Expression In permeabilized Cells Principle Immunoblot Analysis of phox Subunits of nox2 Principle Conclusion Acknowledgement References Chapter 28 Interferon Gamma Release Assays for Tuberculosis d Quantiferon Assay TSPOT.TB Summary References Chapter 29 Methods for evaluating Antigen-specific Cytotoxic T Lymphocyte Frequency and activity Introduction Note On assay Optimization Functional Assays Target Cell Elimination Detection of granzyme-b Delivery In target Cells (grantoxilux Assay) Summary and limitations Quantitative Assays Intracellular Cytokine Staining (ics) Degranulation Calcium Flux Summary and limitations References Section F Cytokines Chapter 30 Introduction References Chapter 31 Overview of the Cytokine Assay Multiverse: Systems and applications Introduction Cytokine Measurements and multiplex Platforms Overview of the Cytokine Multiverse: Applications and system Assays Cytokine Blockade in Rheumatoid Arthritis (ra) and Systemic Lupus Erythematous (sle) Cytokine Il-6 and multibiomarker Disease Activity Multiplex Cytokine Assays and covid-19 Beyond Measurement: Cytokine Signaling Conversion As an Immersive Immunotherapy Tool Summary Acknowledgements References Chapter 32 Monitoring Interferons In autoimmune Diseases and interferonopathies Introduction Interferons In the Etiology of autoimmune Diseases and interferonopathies Autoimmune Diseases Interferonopathies IFNs In diagnostics Interferons in the Diagnosis of Sle Interferons In the Diagnosis of interferonopathies Methods for ifn Measurement With potential Diagnostic Application Direct IFN Measurement Measurement of IFN-stimulated Transcripts References Chapter 33 Detection of anti-cytokine Autoantibodies and clinical Applications Detection Strategies Enzyme-linked Immunosorbent Assay (elisa) Bead-based Immunoassays Luciferase Immunoprecipitation Systems (lips) Radioimmunoassays Immunoblotting Protein Arrays Functional Assays Isotype and subclass Analysis Titer Mechanisms of action Other Diagnostic Considerations Summary Acknowledgments References Section G Laboratory Management Chapter 34 Licensure and accreditation Introduction CLIA State Laboratory Licensing Laboratory Accreditation Clia Mandate Personnel Classifications and requirements Chapter 35 Validation and Quality Control: General Principles and Application to the Clinical Immunology Laboratory Introduction Guidelines for assay Development and validation Classification of assays FDA-approved, Unmodified Assays FDA-approved, Modified Assays Laboratory-developed Tests Classification According To performance Characteristics Qualitative Assays Quasi-quantitative Assays Semiquantitative or Relative Quantitative Assays Quantitative Assays Development and validation Prior To validation Validation Parameters Accuracy Precision Lob, Lod, and loq Linearity Reference Range or Reference Interval Analytical Sensitivity and specificity Clinical (diagnostic) Sensitivity Clinical (diagnostic) Specificity Quality Control Quality Control Material Tools To monitor Quality Levey Jennings Charts Internal Controls Delta Checks Results Trending Automated Liquid Handling Systems Concluding Remarks References Chapter 36 Quality Assurance and Quality Management of the Medical Laboratory Introduction Quality Assurance Phases of clinical Testing: Preanalytic, Analytic, Postanalytic Individualized QC Plan (iqcp) Role of proficiency Testing (pt)/EQAs Delta Check, Biological Variation (bv) and the Reference Change Value (rcv) New Qm Initiative In the Medical Lab: Role of the Diagnostic Management Team (dmt) The Choosing Wisely Campaign Concluding Discussion Points References Section H Infectious Diseases Caused by Bacteria, Mycoplasmas, Chlamydiae, and Rickettsiae Chapter 37 Introduction Chapter 38 Diagnostic Methods for Group a Streptococcal Infections Introduction Clinical Description and epidemiology Pathogenesis of arf and agn Antibody Responses to Group a Streptococcus Diagnostic Methods Rapid Streptococcal Antigen Detection of antibodies To extracellular Antigens Aso Test Nephelometric Method Anti-dnase B Test Research-use-only Tests Antihyaluronidase Test Antistreptokinase Test Anti-nadase Test Detection of antibodies To cellular Antigens Anti-c5a Peptidase Antibodies (anti-scpa) Anti-a Carbohydrate Test Anti-m Protein Antibodies Anti-spna Antibodies Conclusions References Chapter 39 Laboratory Diagnosis of Syphilis Introduction Clinical Features Epidemiology Diagnosis Laboratory Testing Indications Methods Direct Detection Serology Nontreponemal Tests Treponemal Antibody Tests Rapid Point-of-care Tests Quality Control and assurance Syphilis Testing Algorithms Performance Characteristics Limitations Interpretation References Chapter 40 Lyme Disease, Relapsing Fever, and Leptospirosis Introduction Lyme Borreliosis Taxonomy Epidemiology Clinical Description Diagnosis Laboratory Testing Relapsing Fever Taxonomy Epidemiology Clinical Description Diagnosis and laboratory Testing Leptospirosis Taxonomy Epidemiology Clinical Description Diagnosis Laboratory Testing References Chapter 41 Immunological Tests In tuberculosis Introduction Clinical Description Diagnosis Screening Tests for latent Tb Infection Tuberculin Testing IFN- Release Assays Serologic Tests for active Tb Infection Urine Antigen Testing for the Diagnosis of active tb infection References Chapter 42 Mycoplasma: : Serological Methods Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Indications Available Methods Performance Characteristics Limitations Interpretation References Chapter 43 Chlamydia Infections Introduction Chlmaydia Trachomatis Clinical Description Epidemiology Diagnosis Laboratory Testing Indications Available Methods Performance Characteristics Limitations and interpretation Chlamydia Pneumoniae Clinical Description Epidemiology Diagnosis Laboratory Testing Chlamydia Psittaci Clinical Description Epidemiology Diagnosis Laboratory Testing References Chapter 44 the Rickettsiaceae, Anaplasmataceae, and Coxiellaceae Taxonomy Clinical Description Spotted Fever and typhus Group Rickettsioses Scrub Typhus Ehrlichiosis and anaplasmosis Neorickettsiosis and neoehrlichiosis Q Fever Epidemiology Spotted Fever and typhus Group Rickettsioses Scrub Typhus Ehrlichiosis and anaplasmosis Neorickettsiosis and neoehrlichiosis Q Fever Diagnosis Laboratory Testing Serology Immunohistochemistry Interpretation References Chapter 45 the Bartonellaceae, Brucellaceae, and francisellaceae Introduction Taxonomy Bartonella Brucella Francisella Epidemiology Bartonella Brucella Francisella Clinical Manifestations Bartonella Brucella Francisella Laboratorydiagnosis Bartonella Brucella Francisella References Section I Mycotic and parasitic Diseases Chapter 46 Introduction Chapter 47 Immunological and molecular Approaches for the diagnosis of parasitic Infections Introduction Blood and tissue Parasites Protozoa Nematodes Trematodes Filaria Cestodes Gastrointestinal Parasites Protozoa Nematodes References Chapter 48 Serological Diagnosis of fungal Infections Introduction Methods Used for the Diagnosis of fungal Infections Immunodiffusion Commercial Id Test Procedures Complement Fixation Assay Serodiagnosis of fungal Infections Aspergillosis Blastomycosis Candidiasis Coccidioidomycosis Cryptococcosis Histoplasmosis Paracoccidioidomycosis Talaromyces Marneffei Infection Pneumocystosis Sporotrichosis Suppliers Acknowledgments References Section J Viral Diseases Chapter 49 Introduction Chapter 50 Herpes Simplex Virus and varicella-zoster Virus Introduction Hsv Clinical Description Primary and nonprimary Infection Reactivation Complications Neonatal Herpes Hsv Epidemiology Hsv Diagnosis Hsv Laboratory Testing Indications Available Methods Performance Characteristics Limitations Interpretation Vzv Clinical Description Primary Infection Recurrent Disease Congenital Varicella Syndrome and Neonatal varicella Vzv Epidemiology Vzv Diagnosis Vzv Laboratory Testing Indications Available Methods Performance Characteristics Limitations Interpretations References Chapter 51 Epstein-barr Virus and cytomegalovirus Epstein-barr Virus (ebv) Background and biology Indications for laboratory Tests and their Interpretation Laboratory Methods Cytomegalovirus (cmv) Background and biology Clinical Relevance Laboratory Methods Performance Characteristics Limitations References Chapter 52 Human Herpesviruses 6a, 6b, 7, and 8 HHV-6a/b Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing HHV-7 Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing HHV-8 Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Acknowledgement References Chapter 53 Parvovirus B19 Introduction Clinical Description Erythema Infectiosum Arthropathy Transient Aplastic Crisis Pure Red Cell Aplasia Fetal Infection Other Disease Associations Epidemiology Diagnosis Laboratory Testing Indications Available Methods Performance Characteristics Limitations Interpretation References Chapter 54 Severe Acute Respiratory Syndrome Coronavirus-2 Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Indications Available Methods Sars-cov-2 molecular/naat Assays Genotyping Sars-cov-2 Assays Performance Characteristics Limitations Interpretation References Chapter 55 Enteroviruses Introduction Clinical Description Severe Respiratory Disease Myocarditis Hand-foot-mouth Disease Acute Hemorrhagic Conjunctivitis Neurological Illness Epidemiology Diagnosis Laboratory Testing Indications Available Methods Performance Characteristics Interpretation Limitations References Chapter 56 Measles, Mumps, and rubella Viruses Introduction Clinical Description Measles Mumps Rubella Epidemiology Measles Mumps Rubella Diagnosis Laboratory Testing Indications Available Methods Performance Characteristics Limitations Interpretation References Chapter 57 Viral Hepatitis Introduction Hepatitis A virus Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Hepatitis B Virus Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Hepatitis C Virus Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Hepatitis Delta Virus Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Hepatitis E Virus Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing References Chapter 58 Arboviruses Introduction Clinical Presentation Generalized Febrile Illness Hemorrhagic Fever Neuroinvasive Disease Epidemiology Diagnosis Laboratory Testing Indications Available Methods Performance Characteristics Limitations Interpretation References Chapter 59 Hantavirus Infections Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Indications Available Methods Molecular Tests Performance Characteristics Limitations Interpretation References Chapter 60 Rabies Virus Introduction Clinical Description Epidemiology Diagnosis Considerations for working With rabv Laboratory Testing Indications Available Methods Acknowledgments References Chapter 61 Human T-cell Lymphotropic Virus Types 1 and 2 Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Indications Available Methods Molecular Diagnosis Performance Characteristics Limitations Interpretation References Chapter 62 Human Immunodeficiency Virus Introduction Clinical Description Epidemiology Diagnosis Laboratory Testing Indications Available Methods Performance Characteristics Limitations Interpretation References Chapter 63 Bandavirus Introduction Clinical Description Epidemiology Heartland Virus Severe Fever With thrombocytopenia Syndrome virus Bhanja Virus Diagnosis Laboratory Testing Indications Available Methods Performance Characteristics Limitations Interpretation References Volume 2 Title Page Coyright Page Contents Editorial Board Contributors Preface Author and Editor Conflicts of Interest Section K Errors ofimmunity Chapter 64 Introduction to inborn Errors of Immunity References Chapter 65 Newborn Screening for Severe Combined Immunodeficiency Overview Impact of SCID Newborn Screening (purpose, Benefit) Methodology ImmunologIcal Background of TREC-based SCID NBS Commercial Assays and in-house Assays Some Considerations For interpretation Follow-up of abnormal TREC NBS Confirmatory Testing Physiologic T Cell Lymphopenia Due to Prematurity Diagnostic Testing SCID Non-SCID Causes of abnormal TREC NBS Secondary Conditions Treatment Future Directions ADA Deficiency Krec Detection of 22q11.2 Copy Number in 22q11.2 Deletion Syndrome Proteomics Conclusion References Chapter 66 Combined Immunodeficiencies Introduction Diagnostic Methodologies Immunophenotyping Functional Testing T-Cell Diversity/repertoire Radiosensitivity Genetic Testing ClinIcal Examples Severe Combined Immune Deficiency Treatment Conclusions References Chapter 67 Predominantly Antibody Deficiencies Disorders ChaRACterized by Predominantly Antibody Deficiencies Who Should Be investIgAted For predominantly Antibody Deficiencies: ClinIcal Manifestations of predominantly Antibody Deficiencies Key Concepts Major Categories of predominantly Antibody Deficiencies Antibody Deficiencies With absent B Cells Common Variable Immune Deficiency Antibody Deficiency Due to A defect in immunoglobulin Isotope Switching IgA Deficiency IgG Subclass Deficiency IgM Deficiency Specific Antibody Deficiency Transient Hypogammaglobulinemia of infancy Approach To evaluation of patients With suspected Predominantly Antibody Deficiencies The Laboratory investIgAtion of Predominantly Antibody Deficiency Methods Treatment Genetic ANAlysis Conclusions Acknowledgments References Chapter 68 Primary Immune Regulatory Disorders Introduction Immunopathophysiology and clinIcal Clusters of pird Organ-specific Manifestations General Methodology: Pird Diagnosis and testing Genetic Diagnosis of pirds Laboratory Testing For pirds ClinIcal Examples: Key Monogenic Pirds IPex/IPex-like Tregopathies Pi-3 kinase (pi3k) Gene Defects: Activated Pi3k Syndrome (apds) and inactivated Pi3k Syndrome (IPgs) caRD11/cbm Complex Defects Future Directions References Chapter 69 Primary innate Immune Defects (including Phagocytic Disorders, TLR Defects, and NK Cell Deficiencies)