Laboratory Manual
, Contents
Editorial Board / xi 6 Immunoglobulin Genes / 51
THOMAS J. KIPPS, EMANUELA M. GHIA, AND
Contributors / xiii LAURA Z. RASSENTI
Foreword: How It Began / xxiii 7 Immunoglobulin Quantification and Viscosity
Measurement / 65
Preface / xxv
JEFFREY S. WARREN
Author and Editor Conflicts of Interest / xxvii 8 Clinical Indications and Applications of Serum
and Urine Protein Electrophoresis / 74
DAVID F. KEREN AND RICHARD L. HUMPHREY
s ec tion A 9 Immunochemical Characterization of
Immunoglobulins in Serum, Urine, and
Cerebrospinal Fluid / 89
General Methods / 1 ELIZABETH SYKES AND YVONNE POSEY
VOLUME EDITOR: ROBERT G. HAMILTON 10 Cryoglobulins, Cryofibrinogenemia, and
SECTION EDITOR: THOMAS A. FLEISHER Pyroglobulins / 101
PETER D. GOREVIC AND DENNIS GALANAKIS
1 Introduction / 3
THOMAS A. FLEISHER 11 Strategy for Detecting and Following
Monoclonal Gammopathies / 112
2 Molecular Methods for Diagnosis of Genetic JERRY A. KATZMANN AND DAVID F. KEREN
Diseases Involving the Immune System / 5
AMY P. HSU
3 The Human Microbiome and Clinical
Immunology / 19
FREDERIC D. BUSHMAN
s ec tion C
4 Protein Analysis in the Clinical Immunology Complement / 125
Laboratory / 26
VOLUME EDITOR: ROBERT G. HAMILTON
ROSHINI SARAH ABRAHAM AND
DAVID R. BARNIDGE SECTION EDITOR: PATRICIA C. GICLAS
12 Introduction / 127
PATRICIA C. GICLAS
s ec tion B 13 The Classical Pathway of Complement / 129
PATRICIA C. GICLAS
14 Analysis of Activity of Mannan-Binding Lectin,
Immunoglobulin an Initiator of the Lectin Pathway of the
Methods =/ 47 Complement System / 133
STEFFEN THIEL
VOLUME EDITOR: ROBERT G. HAMILTON
15 The Nature of the Diseases That Arise from
SECTION EDITOR: DAVID F. KEREN Improper Regulation of the Alternative Pathway
5 Introduction / 49 of Complement / 138
DAVID F. KEREN RICHARD J. H. SMITH
v
AL G R A WA N Y
,vi ■ CONTENTS
s ec tion D s ec tion E
Flow Cytometry / 145
Functional Cellular
VOLUME EDITOR: JOHN L. SCHMITZ
SECTION EDITOR: MAURICE R. G. O’GORMAN Assays / 259
16 Introduction / 147 VOLUME EDITOR: BARBARA DETRICK
MAURICE R. G. O’GORMAN SECTION EDITOR: STEVEN D. DOUGLAS
17 Polychromatic Flow Cytometry / 149 26 Introduction / 261
ANGÉLIQUE BIANCOTTO AND STEVEN D. DOUGLAS
J. PHILIP McCOY, JR.
27 Cryopreservation of Peripheral Blood
18 High-Sensitivity Detection of Red and Mononuclear Cells / 263
White Blood Cells in Paroxysmal Nocturnal ADRIANA WEINBERG
Hemoglobinuria by Multiparameter Flow
Cytometry / 168 28 Lymphocyte Activation / 269
ANDREA ILLINGWORTH, MICHAEL KEENEY, ROSHINI SARAH ABRAHAM
AND D. ROBERT SUTHERLAND 29 Functional Assays for B Cells and
19 Standardized Flow Cytometry Assays for Antibodies / 280
Enumerating CD34+ Hematopoietic Stem MOON H. NAHM AND ROBERT L. BURTON
Cells / 182 30 Methods for Detection of Antigen-Specific
D. ROBERT SUTHERLAND AND T Cells by Enzyme-Linked Immunospot Assay
MICHAEL KEENEY (ELISPOT) / 290
20 Functional Flow Cytometry-Based Assays BARBARA L. SHACKLETT AND
of Myeloid and Lymphoid Functions for DOUGLAS F. NIXON
the Diagnostic Screening of Primary 31 Regulatory T Cell (Treg) Assays: Repertoire,
Immunodeficiency Diseases / 199 Functions, and Clinical Importance of Human
MAURICE R. G. O’GORMAN Treg / 296
21 Acute Lymphoblastic Leukemia/Lymphoma: THERESA L. WHITESIDE
Diagnosis and Minimal Residual Disease 32 Measurement of NK Cell Phenotype and Activity
Detection by Flow Cytometric in Humans / 300
Immunophenotyping / 207 SAMUEL C. C. CHIANG AND
JOSEPH A. DiGIUSEPPE YENAN T. BRYCESON
22 Acute Myeloid Leukemia: Diagnosis and 33 Functional Assays for the Diagnosis of Chronic
Minimal Residual Disease Detection by Flow Granulomatous Disease / 310
Cytometry / 217 DEBRA LONG PRIEL AND DOUGLAS B. KUHNS
BRENT WOOD AND LORI SOMA
23 Chronic Lymphocytic Leukemia, the Prototypic
Chronic Leukemia for Flow Cytometric
Analysis / 226
HEBA DEGHEIDY, DALIA A. A. SALEM,
s ec tion F
CONSTANCE M. YUAN, AND Cytokines and
MARYALICE STETLER-STEVENSON
Chemokines / 321
24 Plasma Cell Disorders / 235
JUAN FLORES-MONTERO, LUZALBA SANOJA, VOLUME EDITOR: BARBARA DETRICK
JOSÉ JUAN PÉREZ, FANNY POJERO, SECTION EDITOR: JOHN J. HOOKS
NOEMÍ PUIG, MARÍA BELÉN VIDRIALES, AND
ALBERTO ORFAO 34 Introduction / 323
JOHN J. HOOKS
25 Future Cytometric Technologies and
Applications / 251 35 Multiplex Cytokine Assays / 324
HOLDEN T. MAECKER ELIZABETH R. DUFFY AND DANIEL G. REMICK
36 Cytokine Measurement by Flow
Cytometry / 338
HOLDEN T. MAECKER
37 Chemokine and Chemokine Receptor
Analysis / 343
SABINA A. ISLAM, BENJAMIN D. MEDOFF, AND
, CONTENTS ■ vii
ANDREW D. LUSTER 48 Immunological Tests in Tuberculosis / 433
38 Cytokines: Diagnostic and Clinical CHRISTINE M. LITWIN
Applications / 357 49 Mycoplasma: Immunologic and Molecular
PRIYANKA VASHISHT AND TIMOTHY B. NIEWOLD Diagnostic Methods / 444
39 Detection of Anticytokine Autoantibodies and KEN B. WAITES, MARY B. BROWN, AND
Clinical Applications / 365 JERRY W. SIMECKA
SARAH K. BROWNE 50 Chlamydia and Chlamydophila
Infections / 453
ROSEMARY SHE
s ec tion G 51 The Rickettsiaceae, Anaplasmataceae, and
Coxiellaceae / 461
LUCAS S. BLANTON AND DAVID H. WALKER
Immunohistology and 52 The Bartonellaceae, Brucellaceae, and
Immunopathology / 373 Francisellaceae / 473
CHRISTINE M. LITWIN, BURT ANDERSON,
VOLUME EDITOR: ROBERT G. HAMILTON RENEE TSOLIS, AND AMY RASLEY
SECTION EDITOR: R. NEAL SMITH
40 Introduction / 375
ROBERT G. HAMILTON
41 Immunofluorescence Methods in the Diagnosis
of Renal and Cardiac Diseases / 376
s ec tion I
A. BERNARD COLLINS, JAMES R. STONE, AND Mycotic and Parasitic
R. NEAL SMITH Diseases / 483
42 Western Blot Analysis for the Detection
VOLUME EDITOR: ROBERT G. HAMILTON
of Anti-Glomerular Basement Membrane
Antibodies and Anti-Phospholipase A2 Receptor SECTION EDITOR: THOMAS B. NUTMAN
Antibodies / 385 53 Introduction / 485
A. BERNARD COLLINS AND R. NEAL SMITH THOMAS B. NUTMAN
54 Immunological and Molecular Approaches for
s ec tion H the Diagnosis of Parasitic Infections / 486
PATRICIA P. WILKINS AND THOMAS B. NUTMAN
55 Serological and Molecular Diagnosis of Fungal
Infectious Diseases Caused Infections / 503
MARK D. LINDSLEY
by Bacteria, Mycoplasmas,
Chlamydiae, and
Rickettsiae / 391
VOLUME EDITOR: JOHN L. SCHMITZ
s ec tion J
SECTION EDITOR: CHRISTINE M. LITWIN Viral Diseases / 535
43 Introduction / 393 VOLUME EDITOR: JOHN L. SCHMITZ
CHRISTINE M. LITWIN SECTION EDITORS: RICHARD L. HODINKA AND
44 Diagnostic Methods for Group A Streptococcal JOHN L. SCHMITZ
Infections / 394 56 Introduction / 537
CHRISTINE M. LITWIN, SHELDON E. LITWIN, JOHN L. SCHMITZ
AND HARRY R. HILL
57 Immunologic and Molecular Methods for Viral
45 Diagnosis of Helicobacter pylori Infection and Diagnosis / 538
Assessment of Eradication / 404 MARIE LOUISE LANDRY AND YI-WEI TANG
BRUCE E. DUNN AND SUHAS H. PHADNIS
58 Herpes Simplex Virus / 550
46 Laboratory Diagnosis of Syphilis / 412 D. SCOTT SCHMID
JOHN L. SCHMITZ
59 Varicella-Zoster Virus / 556
47 Lyme Disease, Relapsing Fever, and
Leptospirosis / 419 D. SCOTT SCHMID
GUIQING WANG AND 60 Epstein-Barr Virus and Cytomegalovirus / 563
MARIA E. AGUERO-ROSENFELD HENRY H. BALFOUR, JR., KRISTIN A. HOGQUIST,
AND PRIYA S. VERGHESE
AL G R A WA N Y
, Contents
Editorial Board / xi 6 Immunoglobulin Genes / 51
THOMAS J. KIPPS, EMANUELA M. GHIA, AND
Contributors / xiii LAURA Z. RASSENTI
Foreword: How It Began / xxiii 7 Immunoglobulin Quantification and Viscosity
Measurement / 65
Preface / xxv
JEFFREY S. WARREN
Author and Editor Conflicts of Interest / xxvii 8 Clinical Indications and Applications of Serum
and Urine Protein Electrophoresis / 74
DAVID F. KEREN AND RICHARD L. HUMPHREY
s ec tion A 9 Immunochemical Characterization of
Immunoglobulins in Serum, Urine, and
Cerebrospinal Fluid / 89
General Methods / 1 ELIZABETH SYKES AND YVONNE POSEY
VOLUME EDITOR: ROBERT G. HAMILTON 10 Cryoglobulins, Cryofibrinogenemia, and
SECTION EDITOR: THOMAS A. FLEISHER Pyroglobulins / 101
PETER D. GOREVIC AND DENNIS GALANAKIS
1 Introduction / 3
THOMAS A. FLEISHER 11 Strategy for Detecting and Following
Monoclonal Gammopathies / 112
2 Molecular Methods for Diagnosis of Genetic JERRY A. KATZMANN AND DAVID F. KEREN
Diseases Involving the Immune System / 5
AMY P. HSU
3 The Human Microbiome and Clinical
Immunology / 19
FREDERIC D. BUSHMAN
s ec tion C
4 Protein Analysis in the Clinical Immunology Complement / 125
Laboratory / 26
VOLUME EDITOR: ROBERT G. HAMILTON
ROSHINI SARAH ABRAHAM AND
DAVID R. BARNIDGE SECTION EDITOR: PATRICIA C. GICLAS
12 Introduction / 127
PATRICIA C. GICLAS
s ec tion B 13 The Classical Pathway of Complement / 129
PATRICIA C. GICLAS
14 Analysis of Activity of Mannan-Binding Lectin,
Immunoglobulin an Initiator of the Lectin Pathway of the
Methods =/ 47 Complement System / 133
STEFFEN THIEL
VOLUME EDITOR: ROBERT G. HAMILTON
15 The Nature of the Diseases That Arise from
SECTION EDITOR: DAVID F. KEREN Improper Regulation of the Alternative Pathway
5 Introduction / 49 of Complement / 138
DAVID F. KEREN RICHARD J. H. SMITH
v
AL G R A WA N Y
,vi ■ CONTENTS
s ec tion D s ec tion E
Flow Cytometry / 145
Functional Cellular
VOLUME EDITOR: JOHN L. SCHMITZ
SECTION EDITOR: MAURICE R. G. O’GORMAN Assays / 259
16 Introduction / 147 VOLUME EDITOR: BARBARA DETRICK
MAURICE R. G. O’GORMAN SECTION EDITOR: STEVEN D. DOUGLAS
17 Polychromatic Flow Cytometry / 149 26 Introduction / 261
ANGÉLIQUE BIANCOTTO AND STEVEN D. DOUGLAS
J. PHILIP McCOY, JR.
27 Cryopreservation of Peripheral Blood
18 High-Sensitivity Detection of Red and Mononuclear Cells / 263
White Blood Cells in Paroxysmal Nocturnal ADRIANA WEINBERG
Hemoglobinuria by Multiparameter Flow
Cytometry / 168 28 Lymphocyte Activation / 269
ANDREA ILLINGWORTH, MICHAEL KEENEY, ROSHINI SARAH ABRAHAM
AND D. ROBERT SUTHERLAND 29 Functional Assays for B Cells and
19 Standardized Flow Cytometry Assays for Antibodies / 280
Enumerating CD34+ Hematopoietic Stem MOON H. NAHM AND ROBERT L. BURTON
Cells / 182 30 Methods for Detection of Antigen-Specific
D. ROBERT SUTHERLAND AND T Cells by Enzyme-Linked Immunospot Assay
MICHAEL KEENEY (ELISPOT) / 290
20 Functional Flow Cytometry-Based Assays BARBARA L. SHACKLETT AND
of Myeloid and Lymphoid Functions for DOUGLAS F. NIXON
the Diagnostic Screening of Primary 31 Regulatory T Cell (Treg) Assays: Repertoire,
Immunodeficiency Diseases / 199 Functions, and Clinical Importance of Human
MAURICE R. G. O’GORMAN Treg / 296
21 Acute Lymphoblastic Leukemia/Lymphoma: THERESA L. WHITESIDE
Diagnosis and Minimal Residual Disease 32 Measurement of NK Cell Phenotype and Activity
Detection by Flow Cytometric in Humans / 300
Immunophenotyping / 207 SAMUEL C. C. CHIANG AND
JOSEPH A. DiGIUSEPPE YENAN T. BRYCESON
22 Acute Myeloid Leukemia: Diagnosis and 33 Functional Assays for the Diagnosis of Chronic
Minimal Residual Disease Detection by Flow Granulomatous Disease / 310
Cytometry / 217 DEBRA LONG PRIEL AND DOUGLAS B. KUHNS
BRENT WOOD AND LORI SOMA
23 Chronic Lymphocytic Leukemia, the Prototypic
Chronic Leukemia for Flow Cytometric
Analysis / 226
HEBA DEGHEIDY, DALIA A. A. SALEM,
s ec tion F
CONSTANCE M. YUAN, AND Cytokines and
MARYALICE STETLER-STEVENSON
Chemokines / 321
24 Plasma Cell Disorders / 235
JUAN FLORES-MONTERO, LUZALBA SANOJA, VOLUME EDITOR: BARBARA DETRICK
JOSÉ JUAN PÉREZ, FANNY POJERO, SECTION EDITOR: JOHN J. HOOKS
NOEMÍ PUIG, MARÍA BELÉN VIDRIALES, AND
ALBERTO ORFAO 34 Introduction / 323
JOHN J. HOOKS
25 Future Cytometric Technologies and
Applications / 251 35 Multiplex Cytokine Assays / 324
HOLDEN T. MAECKER ELIZABETH R. DUFFY AND DANIEL G. REMICK
36 Cytokine Measurement by Flow
Cytometry / 338
HOLDEN T. MAECKER
37 Chemokine and Chemokine Receptor
Analysis / 343
SABINA A. ISLAM, BENJAMIN D. MEDOFF, AND
, CONTENTS ■ vii
ANDREW D. LUSTER 48 Immunological Tests in Tuberculosis / 433
38 Cytokines: Diagnostic and Clinical CHRISTINE M. LITWIN
Applications / 357 49 Mycoplasma: Immunologic and Molecular
PRIYANKA VASHISHT AND TIMOTHY B. NIEWOLD Diagnostic Methods / 444
39 Detection of Anticytokine Autoantibodies and KEN B. WAITES, MARY B. BROWN, AND
Clinical Applications / 365 JERRY W. SIMECKA
SARAH K. BROWNE 50 Chlamydia and Chlamydophila
Infections / 453
ROSEMARY SHE
s ec tion G 51 The Rickettsiaceae, Anaplasmataceae, and
Coxiellaceae / 461
LUCAS S. BLANTON AND DAVID H. WALKER
Immunohistology and 52 The Bartonellaceae, Brucellaceae, and
Immunopathology / 373 Francisellaceae / 473
CHRISTINE M. LITWIN, BURT ANDERSON,
VOLUME EDITOR: ROBERT G. HAMILTON RENEE TSOLIS, AND AMY RASLEY
SECTION EDITOR: R. NEAL SMITH
40 Introduction / 375
ROBERT G. HAMILTON
41 Immunofluorescence Methods in the Diagnosis
of Renal and Cardiac Diseases / 376
s ec tion I
A. BERNARD COLLINS, JAMES R. STONE, AND Mycotic and Parasitic
R. NEAL SMITH Diseases / 483
42 Western Blot Analysis for the Detection
VOLUME EDITOR: ROBERT G. HAMILTON
of Anti-Glomerular Basement Membrane
Antibodies and Anti-Phospholipase A2 Receptor SECTION EDITOR: THOMAS B. NUTMAN
Antibodies / 385 53 Introduction / 485
A. BERNARD COLLINS AND R. NEAL SMITH THOMAS B. NUTMAN
54 Immunological and Molecular Approaches for
s ec tion H the Diagnosis of Parasitic Infections / 486
PATRICIA P. WILKINS AND THOMAS B. NUTMAN
55 Serological and Molecular Diagnosis of Fungal
Infectious Diseases Caused Infections / 503
MARK D. LINDSLEY
by Bacteria, Mycoplasmas,
Chlamydiae, and
Rickettsiae / 391
VOLUME EDITOR: JOHN L. SCHMITZ
s ec tion J
SECTION EDITOR: CHRISTINE M. LITWIN Viral Diseases / 535
43 Introduction / 393 VOLUME EDITOR: JOHN L. SCHMITZ
CHRISTINE M. LITWIN SECTION EDITORS: RICHARD L. HODINKA AND
44 Diagnostic Methods for Group A Streptococcal JOHN L. SCHMITZ
Infections / 394 56 Introduction / 537
CHRISTINE M. LITWIN, SHELDON E. LITWIN, JOHN L. SCHMITZ
AND HARRY R. HILL
57 Immunologic and Molecular Methods for Viral
45 Diagnosis of Helicobacter pylori Infection and Diagnosis / 538
Assessment of Eradication / 404 MARIE LOUISE LANDRY AND YI-WEI TANG
BRUCE E. DUNN AND SUHAS H. PHADNIS
58 Herpes Simplex Virus / 550
46 Laboratory Diagnosis of Syphilis / 412 D. SCOTT SCHMID
JOHN L. SCHMITZ
59 Varicella-Zoster Virus / 556
47 Lyme Disease, Relapsing Fever, and
Leptospirosis / 419 D. SCOTT SCHMID
GUIQING WANG AND 60 Epstein-Barr Virus and Cytomegalovirus / 563
MARIA E. AGUERO-ROSENFELD HENRY H. BALFOUR, JR., KRISTIN A. HOGQUIST,
AND PRIYA S. VERGHESE
AL G R A WA N Y
