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.
Table of Contents
Volume 1
Editorial Board xi
Contributors xiii
Preface xxiii
Author and Editor Conflicts of Interest xxv
Section A
Molecular Methods 1
Volume Editor: John L. Schmitz
Section Editor: Eric T. Weimer
1 Introduction 3
Eric T. Weimer
2 Molecular Methods for Genetic Diagnosis of Inborn Errors of Immunity 5
Sarah L. Anzick and Amy P. Hsu
3 Immunoglobulin and T- Cell Receptor Gene Assessment 23
Paul A. Wadsworth, Chandler C. Ho, and Bing M. Zhang
Section B
Immunoglobulin Methods 39
Volume Editor: Barbara Detrick
Section Editor: David F. Keren
4 Introduction 41
David F. Keren
5 Clinical Indications and Applications of Serum and Urine Protein Electrophoresis 43
David M. Manthei and David F. Keren
6 Immunochemical Characterization of Immunoglobulins in Serum, Urine, and Cerebrospinal Fluid 62
Caitlin O. Schein and William G. Finn
7 Cryoglobulins and Cryofibrinogenemia 79
Peter D. Gorevic, Dennis Galanakis, and Jeffrey S. Warren
8 Strategy for Detecting and Following Monoclonal Gammopathies 90
David F. Keren and Lee Schroeder
Section C
Complement 107
Volume Editor: Barbara Detrick
Section Editor: Ashley Frazer-Abel
9 Introduction 109
Ashley Frazer- Abel
10 Methods of Complement Analysis 111
Diana Pauly and Michael Kirschfink
11 Assessing Complement Dysregulation in the Thrombotic Microangiopathies and Ultrarare Complement- Mediated Kidney Disease 119
Amanda K. Slagle, Jill J. Hauer, and Richard J. H. Smith
12 Current and Future of Complement Therapeutics and their Impact on the Clinical Immunology Laboratory 129
Bradley Dixon, Shrey Purohit, and Ashley Frazer- Abel
Section D
Flow Cytometry 139
Volume Editor: Maurice R. G. O’gorman
Section Editor: Paul K. Wallace
13 Introduction: Clinical Flow Cytometry Applications and Quality Control 141
Paul K. Wallace
14 Polychromatic Flow Cytometry 144
Ankit Saxena, Angelique Biancotto, Pradeep Dagur, And John Philip Mccoy, Jr
15 High- Sensitivity Detection of Paroxysmal Nocturnal Hemoglobinuria by Multiparameter Flow Cytometry 166
Ahmad Al- Attar, Stephen J. Richards, Iuri Marinov, Andrea Illingworth, And D. Robert Sutherland
16 Standardized Flow Cytometry Assays for Enumerating CD34 + Hematopoietic Stem Cells 189
Ahmad Al- Attar and D. Robert Sutherland
17 The Detection of Fetal Red Cells in Fetomaternal Hemorrhage by Flow Cytometry 213
Bruce Greig
18 Functional Flow Cytometry- Based Assays of Myeloid and Lymphoid Functions for the Diagnostic Screening of Inborn Errors of Immunity 221
Caleb Cornaby and Maurice R. G. O’gorman
19 Quality Assurance and Validation in Clinical Flow Cytometry 231
Teri Oldaker and Antony Bakke
20 Future Cytometric Technologies and Applications 249
Holden T. Maecker
Section E
Functional Cellular Assays 253
Volume Editor: Maurice R. G. O’gorman
Section Editors: Miranda B. Carper And Thomas N. Denny
21 Introduction 255
Miranda B. Carper and Thomas N. Denny
22 Cryopreservation of Blood Mononuclear Cells 257
Cassandra Porth, Ambrosia Garcia, Raul Louzao, Krystle Garth, Emilie Jalbert, and Adriana Weinberg
23 Functional Assays for B Cells and Antibodies 266
David C. Lafon and Moon H. Nahm
24 Methods for Detection of Antigen- Specific T Cells by Enzyme- Linked Immunospot Assay 277
Mark Berrong, Danielle Nettere, and Guido Ferrari
25 Regulatory T Cell (Treg) Characterization: Phenotype, Function, and Clinical Importance of Human Treg 284
Alma- Martina Cepika and Rosa Bacchetta
26 Measurement of Natural Killer Cell Phenotype and Activity in Humans 292
Heinrich Schlums, Tessa M. Campbell, and Yenan T. Bryceson
27 Functional Assays for the Diagnosis of Chronic Granulomatous Disease 304
Debra Long Priel and Douglas B. Kuhns
28 Interferon Gamma Release Assays for Tuberculosis Diagnosis 318
Sofia Zavala and Jason E. Stout
29 Methods for Evaluating Antigen- Specific Cytotoxic T Lymphocyte Frequency and Activity 324
Danielle Nettere and Guido Ferrari
Section F
Cytokines 333
Volume Editor: Barbara Detrick
Section Editors: John J. Hooks And Timothy B. Niewold
30 Introduction 335
John J. Hooks and Timothy B. Niewold
31 Overview of the Cytokine Assay Multiverse: Systems and Applications 337
Julio C. Valencia, Ngoc Thuy T. Nguyen, And Howard A. Young
32 Monitoring Interferons in Autoimmune Diseases and Interferonopathies 344
Andrra Nimoni and Timothy B. Niewold
33 Detection of Anti-Cytokine Autoantibodies and Clinical Applications 352
Serena J. Lee, Eszter Toth, Lindsey B. Rosen, Sarah K. Browne, and Steven M. Holland
Section G
Laboratory Management 363
Volume Editor: John L. Schmitz
Section Editor: Thomas S. Alexander
34 Licensure and Accreditation 365
Thomas S. Alexander
35 Validation and Quality Control: General Principles and Application to the Clinical Immunology Laboratory 369
Terri Lebo, Patricia A. Merkel, and Vijaya Knight
36 Quality Assurance and Quality Management of the Medical Laboratory 384
Paul R. Johnson
Section H
Infectious Diseases Caused By Bacteria, Mycoplasmas, Chlamydiae, and Rickettsiae 393
Volume Editor: John L. Schmitz
Section Editor: Christine M. Litwin
37 Introduction 395
Christine M. Litwin
38 Diagnostic Methods for Group A Streptococcal Infections 396
Christine M. Litwin, Sheldon E. Litwin, And Harry R. Hill
39 Laboratory Diagnosis of Syphilis 404
Tyson Chiaro and John L. Schmitz
40 Lyme Disease, Relapsing Fever, and Leptospirosis 412
Guiqing Wang and Maria E. Aguero- Rosenfeld
41 Immunological Tests in Tuberculosis 424
Christine M. Litwin
42 Mycoplasma: Serological Methods 433
Ken B. Waites, Mary B. Brown, and Jerry W. Simecka
43 Chlamydia Infections 438
Rosemary C. She and Valentina Zanfagnin
44 The Rickettsiaceae, Anaplasmataceae, and Coxiellaceae 447
Lucas S. Blanton and David H. Walker
45 The Bartonellaceae, Brucellaceae, and Francisellaceae 458
Christine M. Litwin, Burt Anderson, Renee Tsolis, and Amy Rasley
Section I
Mycotic and Parasitic Diseases 467
Volume Editor: John L. Schmitz
Section Editor: Elitza S. Theel
46 Introduction 469
Elitza S. Theel
47 Immunological and Molecular Approaches for the Diagnosis of Parasitic Infections 470
Marc R. Couturier and Elitza S. Theel
48 Serological Diagnosis of Fungal Infections 489
Mark D. Lindsley
Section J
Viral Diseases 521
Volume Editor: John L. Schmitz
Section Editor: Jennifer Dien Bard
49 Introduction 523
Jennifer Dien Bard
50 Herpes Simplex Virus and Varicella- Zoster Virus 524
Susan Realegeno
51 Epstein- Barr Virus and Cytomegalovirus 535
Henry H. Balfour, Jr., Kristin A. Hogquist, And Priya S. Verghese
52 Human Herpesviruses 6A, 6B, 7, and 8 550
Shangxin (shaun) Yang
53 Parvovirus B19 563
Rebecca Yee and Jose A. Lucar
54 Severe Acute Respiratory Syndrome Coronavirus- 2 572
Patricia Slev
55 Enteroviruses 583
Bijal A. Parikh and Allison R. Eberly
56 Measles, Mumps, and Rubella Viruses 592
Ryan F. Relich and Elitza S. Theel
57 Viral Hepatitis 600
Kileen L. Shier
58 Arboviruses 616
Mimi R. Precit and Rachael M. Liesman
59 Hantavirus Infections 634
William Marciel De Souza and Luiz Tadeu Moraes Figueiredo
60 Rabies Virus 640
D. Craig Hooper
61 Human T- Cell Lymphotropic Virus Types 1 and 2 649
Laura Gillim and Ajay Grover
62 Human Immunodeficiency Virus 658
S. Michele Owen
63 Bandavirus 668
Amanda E. Calvert and Holly R. Hughes
Volume 2
Section K
Errors Of Immunity 679
Volume Editor: Maurice R. G. O’gorman
Section Editor: Roshini S. Abraham
64 Introduction to Inborn Errors of Immunity 681
Roshini S. Abraham
65 Newborn Screening for Severe Combined Immunodeficiency 683
Lisa Kobrynski and Mirjam Van Der Burg
66 Combined Immunodeficiencies 692
Tamar S. Rubin, Kelli W. Williams, and Monica G. Lawrence
67 Predominantly Antibody Deficiencies 717
Sara Barmettler, Daniel Digiacomo, and Jocelyn R. Farmer
68 Primary Immune Regulatory Disorders 732
Shanmuganathan Chandrakasan, Gulbu Uzel, Carrie L. Lucas, And Andrew L. Snow
69 Primary Innate Immune Defects (Including Phagocytic Disorders, TLR Defects, and NK Cell Deficiencies) 752
Jenna R. E. Bergerson, Alexandra F. Freeman, Emily M. Mace, And Keith A. Sacco
70 Systemic Autoinflammatory Diseases 768
Xiao Peng, Oskar Schnappauf, Adriana A. De Jesus, and Ivona Aksentijevich
71 Complement Defects 796
Ann M. Moyer, Meera Sridharan, and Maria Alice Vieira Willrich
72 Inherited Bone Marrow Failure Syndromes: Focus on Immunodeficiency and Diagnostic Evaluation 813
Abhishek A. Mangaonkar, Alejandro Ferrer, Asma Ali, Mira Kohorst, and Mrinal M. Patnaik
73 Phenocopies of Inborn Errors of Immunity 825
Elena W. Y. Hsieh, Cullen M. Dutmer, and Vijaya Knight
Section L
llergic Diseases 845
Volume Editor: Barbara Detrick
Section Editor: Robert G. Hamilton
74 Introduction to Allergic Diseases 847
Robert G. Hamilton
75 Quantitation and Standardization of Allergens 849
Michael Brad Strader, Robert Zagorski, Ronald L. Rabin, and Jay E. Slater
76 Immunological Methods in the Diagnostic Allergy Laboratory 860
Robert G. Hamilton
77 In Vitro Assay Methods for Measurement of Mediators and Markers of Allergic Inflammation 866
John T. Schroeder
Section M
Systemic Autoimmune Diseases 877
Volume Editor: Barbara Detrick
Section Editor: Westley H. Reeves
78 Introduction 879
Westley H. Reeves
79 Standardization of Fluorescent Antinuclear Antibody Testing 881
Angela Ceribelli, Antonio Tonutti, Luis E. C. Andrade, and Edward K. L. Chan
80 Immunodiagnosis and Laboratory Assessment of Systemic Lupus Erythematosus 891
Westley H. Reeves, Shuhong Han, and Haoyang Zhuang
81 Immunodiagnosis of Autoimmune Myopathies 900
Minoru Satoh, Angela Ceribelli, Tomoko Hasegawa, Shin Tanaka, Michito Hirakata, and Edward K. L. Chan
82 Immunodiagnosis of Scleroderma 913
Yasuhito Hamaguchi and Masataka Kuwana
83 Immunodiagnosis of Rheumatoid Arthritis 925
Johan Rönnelid, Alf Kastbom, and Carl Turesson
84 Antiphospholipid Antibody Syndrome: Clinical Manifestations and Laboratory Diagnosis 935
Pier Luigi Meroni, Caterina Bodio, Claudia Grossi, and Maria Orietta Borghi
85 Immunodiagnosis of Systemic Vasculitis 945
Jan Damoiseaux
86 Immunodiagnosis of Sjögren’s Disease 952
R. Hal Scofield and Martha Tsaliki
87 Diagnostic Testing for Deficiency of Adenosine Deaminase 2 962
Kailey E. Brodeur, Liang Chen, And Pui Y. Lee
88 Future Direction of Laboratory Testing for Autoimmune Diseases 968
Gerson Dierley Keppeke, Renan De Almeida Agustinelli, Diana Maria Landoni, Wilson Melo Cruvinel, Edward K. L. Chan, And Luis Eduardo Coelho Andrade
Section N
Organ-Localized Autoimmune Diseases 983
Volume Editor: Barbara Detrick
Section Editor: Stanley J. Naides
89 Introduction 985
Stanley J. Naides
90 Endocrinopathies: Hashimoto Thyroiditis, Graves Disease, Hypophysitis, Addison Disease, Premature Ovarian Failure, Male Infertility, and Diabetes 987
Amna Kahn, Giuseppe Barbesino, Jonathan Perez, Ziyan Xu, Xiaofan Jia, Fatima Zaman, Liping Yu, and Patrizio Caturegli
91 Myasthenia Gravis 1014
Hans Frykman
92 Autoantibodies to Brain and Spinal Cord 1024
Raquel Ruiz- García, Laura Naranjo, and Francesc Graus
93 Autoantibodies to Glycolipids in Peripheral Neuropathy 1037
John A. Goodfellow and Hugh J. Willison
94 Detection of Antimitochondrial Autoantibodies in Primary Biliary Cholangitis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis 1044
Patrick S. C. Leung, Michael P. Manns, Ross L. Coppel, and M. Eric Gershwin
95 Cardiovascular Diseases 1054
Jin li
96 Use of Laboratory Tests for Patients with Inflammatory Bowel Diseases 1071
Louma B. Rustam, David E. Elliott, and M. Nedim Ince
97 Celiac Disease 1085
Tracey M. Dafonte, Katherine L. Olshan, And Alessio Fasano
98 Primary Membranous Nephropathy 1090
Ajay Grover, Andre Valcour, and Stanley J. Naides
99 Autoantibodies Directed against Erythrocytes in Autoimmune Hemolytic Anemia and Other Anemias 1098
Laura Cooling
100 Immune Thrombocytopenia 1105
Qizhao Li, Johan Rebetz, Drew Provan, Leendert L. Porcelijn, Rick Kapur, Suzanne Hofstede- Van Egmond, Elisabeth Semple, Li Guo, Shuqian Xu, and John W. Semple
101 Monitoring Autoimmune Reactivity within the Retina 1113
John J. Hooks, Renee Liu, Lucia Sobrin, And Barbara Detrick
102 Diagnosis of Autoimmune Blistering Diseases 1120
Mareike Becker and Enno Schmidt
Section O
Cancer 1133
Volume Editor: Maurice R. G. O’gorman
Section Editor: Brent L. Wood
103 Introduction 1135
Brent L. Wood
104 Immunoassay- Based Tumor Marker Measurement: Assays, Applications, Algorithms, and Additional Approaches 1136
Chinwe Madubata and Alex J. Rai
105 Immune Monitoring during Cancer Immunotherapy 1144
Hesham M. Shehata, Pranay Dogra, and Shomyseh Sanjabi
106 Immunophenotyping by Flow Cytometry of Acute Lymphoblastic Leukemia Lymphoma 1168
Karin P. Miller and Alexandra E. Kovach
107 Acute Myeloid Leukemia: Diagnosis and Measurable Residual Disease Detection by Flow Cytometry 1180
Xueyan Chen and Brent L. Wood
108 Chronic Lymphocytic Leukemia: Diagnosis and Minimal Residual Disease Detection 1193
Min Shi, Dragan Jevremovic, Curtis A. Hanson, and Horatiu Olteanu
109 Evaluation of Plasma Cell Neoplasms 1206
Mikhail Roshal, Qi Gao, Shelby Hutcherson, Katie Thoren, Menglei Zhu, And Kazunory Murata
Section P
Transplantation Immunology 1225
Volume Editor: John L. Schmitz
Section Editor: Gerald P. Morris
110 Introduction: Immunogenetics, Histocompatibility Testing, and Transplantation 1227
Gerald P. Morris
111 Human Leukocyte Antigen Biology and Genetics 1229
Bing Melody Zhang and Marcelo Fernandez- Vina
112 Human Leukocyte Antigen Genotyping for Allogeneic Transplantations 1240
Brian F. Duffy, Mei San Tang, and Chang Liu
113 Human Leukocyte Antigen Antibodies 1262
Olga A. Timofeeva and Sandra Rosen- Bronson
114 Non- HLA Antibodies 1280
Carrie L. Butler, Michelle J. Hickey, Rebecca A. Sosa, and Elaine F. Reed
115 ABO Blood Groups and Transplantation 1291
Anne Halpin, Bruce Motyka, and Lori J. West
116 Cellular Immune Responses in Transplantation 1301
Clarkson Crane and Gerald P. Morris
117 Molecular Characterization of Injury and Rejection in Solid Organ Transplant 1312
Meagan Barner and Zahra Kashi
118 Killer Cell Immunoglobulin- Like Receptors (KIR) in Clinical Transplantation 1323
Raja Rajalingam
119 Chimerism Monitoring 1335
Caroline R. Alquist
Index 1341
