Rupprecht / Nagarajan Current Laboratory Techniques in Rabies Diagnosis, Research and Prevention, Volume 2

1. Auflage 2015
ISBN: 978-0-12-802012-8
Verlag: Elsevier Science & Techn.
Format: EPUB
Kopierschutz: Adobe DRM (»Systemvoraussetzungen)

E-Book, Englisch, 384 Seiten

Laboratory Techniques in Rabies Diagnosis, Research and Prevention provides a basic understanding of the current trends in rabies. It establishes a new facility for rabies surveillance, vaccine and antibody manufacturing. It offers clarity about the choice of laboratory methods for diagnosis and virus typing, of systems for producing monoclonal and polyclonal antibodies and of methods for testing potency of vaccines and antibodies. The book covers advancements in the classical methods described as well as recent methods and approaches pertaining to rabies diagnosis and research. - Supplies techniques pertaining to rabies diagnosis and research - Provides an update on the conventional and modern vaccines for rabies prevention - Offers updates on the full length antibodies and antibody fragments for post exposure prophylaxis of rabies - Presents technique descriptions that can be used to be compared to industry protocols to identify and establish potential new techniques

Autoren/Hrsg.

Rupprecht, Charles

Nagarajan, Thirumeni

Inhaltsverzeichnis

1;Front Cover;1
2;Current Laboratory Techniques in Rabies Diagnosis, Research and Prevention;4
3;Copyright Page;5
4;Contents;6
5;Foreword;14
6;List of Contributors;16
7;PART ONE Rabies Diagnosis;20
7.1;Section A Demonstration of Virus;22
7.1.1;1. Demonstration of Lyssaviruses by Electron Microscopy;24
7.1.1.1;1.1 Introduction;24
7.1.1.2;1.2 Materials;25
7.1.1.2.1;1.2.1 Reagents;25
7.1.1.2.2;1.2.2 Equipment;25
7.1.1.2.3;1.2.3 Biological Materials;26
7.1.1.3;1.3 Methods;26
7.1.1.3.1;1.3.1 Fixation;26
7.1.1.3.1.1;1.3.1.1 Central Nervous System Tissue;26
7.1.1.3.1.2;1.3.1.2 Infected Cell Culture;26
7.1.1.3.2;1.3.2 Processing;27
7.1.1.3.2.1;1.3.2.1 Thin Section Technique;27
7.1.1.3.2.2;1.3.2.2 Negative Staining Technique;28
7.1.1.3.3;1.3.3 Interpretation of Results;28
7.1.1.4;1.4 Discussion;28
7.1.1.4.1;1.4.1 Experimental Tips;28
7.1.1.4.2;1.4.2 Precautions;28
7.1.1.4.3;1.4.3 Alternative Methods;29
7.1.1.4.4;1.4.4 Time Considerations;29
7.1.1.4.5;1.4.5 Limitations;30
7.1.1.5;References;30
7.1.2;2. Virus Isolation in Animals: The Mouse Inoculation Test;32
7.1.2.1;2.1 Ethical Considerations;32
7.1.2.2;2.2 Choice of Mice and their Husbandry;33
7.1.2.3;2.3 Preparation of Inoculum;34
7.1.2.4;2.4 Inoculation Procedure;36
7.1.2.5;2.5 Observation;38
7.1.2.6;2.6 Postmortem Examination;39
7.1.2.7;2.7 Troubleshooting;39
7.1.2.8;2.8 Virus Titration;40
7.1.2.9;References;42
7.1.3;3. Virus Isolation in Cell Culture: The Rabies Tissue Culture Infection Test;44
7.1.3.1;3.1 Introduction;44
7.1.3.2;3.2 Materials;46
7.1.3.2.1;3.2.1 Reagents;46
7.1.3.2.2;3.2.2 Equipment;46
7.1.3.2.3;3.2.3 Biological Materials;47
7.1.3.3;3.3 Methods;47
7.1.3.3.1;3.3.1 Maintenance of Neuroblastoma Cell Cultures and Preparation of the Cell Suspension;47
7.1.3.3.2;3.3.2 Preparation of the Inoculum;47
7.1.3.3.3;3.3.3 Inoculation of 8-chamber Lab-Tek® Slides;47
7.1.3.3.4;3.3.4 Fixation of Cells and Virus Detection;48
7.1.3.3.5;3.3.5 Interpretation of Results;48
7.1.3.4;3.4 Discussion;49
7.1.3.5;References;50
7.1.4;4. A Rat Basophilic Leukemia Cell Sensor for the Detection of Rabies Viruses;52
7.1.4.1;4.1 Introduction;53
7.1.4.2;4.2 Materials;53
7.1.4.2.1;4.2.1 Reagents;53
7.1.4.2.2;4.2.2 Equipment;55
7.1.4.3;4.3 Methods;55
7.1.4.3.1;4.3.1 Making Anti-Rabies Virus Single Domain Antibodies;55
7.1.4.3.2;4.3.2 Construction, Expression, and Purification of Single Domain Antibody Fc Fragment;56
7.1.4.3.3;4.3.3 Antigen Recognition and Receptor Binding of Single Domain Antibody Fc Fragment;56
7.1.4.3.4;4.3.4 Preparation of Rat Basophilic Leukemia Cell Sensor;58
7.1.4.3.5;4.3.5 Preparation of Rabies Virus Sample;58
7.1.4.3.6;4.3.6 Detection of Rabies Virus by Rat Basophilic Leukemia Cell Sensor;58
7.1.4.3.7;4.3.7 Interpretation of Results;59
7.1.4.4;4.4 Discussion;59
7.1.4.4.1;4.4.1 Experimental Tips;59
7.1.4.4.2;4.4.2 Critical Parameters and Troubleshooting;59
7.1.4.4.3;4.4.3 Precautions;59
7.1.4.4.4;4.4.4 The Sensitivity of Rat Basophilic Leukemia Sensor for Detection of Rabies Virus;60
7.1.4.4.5;4.4.5 Time Considerations;61
7.1.4.4.6;4.4.6 Limitations;61
7.1.4.4.7;4.4.7 Future Considerations;61
7.1.4.5;Acknowledgments;61
7.1.4.6;References;62
7.2;Section B Demonstration of Viral Subunits and Antigens;64
7.2.1;5. Purification of Rabies Virus and Its Subunits;66
7.2.1.1;5.1 Introduction;66
7.2.1.2;5.2 Materials and Methods;67
7.2.1.2.1;5.2.1 Purification of Rabies Virus Particles;67
7.2.1.2.2;5.2.2 Purification of Rabies Virus Subunits and Proteins;69
7.2.1.2.2.1;5.2.2.1 Isolation of Biologically Active Subunits;69
7.2.1.2.2.2;5.2.2.2 Solubilization and Purification of Glycoprotein Trimers;69
7.2.1.2.2.3;5.2.2.3 Isolation and Purification of a Soluble Rabies Virus Glycoprotein from the Supernatant of Infected Tissue Cultures;70
7.2.1.2.2.4;5.2.2.4 Purification of a Soluble Recombinant-Expressed Rabies Virus Glycoprotein;71
7.2.1.2.2.5;5.2.2.5 Purification of Rabies Virus Nucleoprotein from Infected Cells;71
7.2.1.2.2.6;5.2.2.6 Purification of Recombinant-Expressed Rabies Virus Nucleoprotein;72
7.2.1.2.2.7;5.2.2.7 Isolation of Denatured Rabies Virus Proteins;72
7.2.1.3;5.3 Discussion;73
7.2.1.4;References;74
7.2.2;6. Preparation of Fluorescent Antibody Conjugate in Rabbits;76
7.2.2.1;6.1 Introduction;76
7.2.2.2;6.2 Materials;77
7.2.2.2.1;6.2.1 Reagents;77
7.2.2.2.2;6.2.2 Equipment;78
7.2.2.2.3;6.2.3 Biological Materials;78
7.2.2.2.4;6.2.4 Laboratory Animals;78
7.2.2.3;6.3 Methods;78
7.2.2.3.1;6.3.1 Propagation of Rabies Virus and Concentration of Ribonucleoprotein;78
7.2.2.3.2;6.3.2 Immunization of Animals and Purification of Immune Globulin G;79
7.2.2.3.3;6.3.3 Conjugation of Immune Globulin to Fluorescein Isothiocyanate;81
7.2.2.3.4;6.3.4 Interpretation of Results;81
7.2.2.4;6.4 Discussion;82
7.2.2.4.1;6.4.1 Critical Parameters and Troubleshooting;83
7.2.2.4.2;6.4.2 Precautions;84
7.2.2.4.3;6.4.3 Alternative Materials and/or Methods;84
7.2.2.4.4;6.4.4 Time Considerations;84
7.2.2.4.5;6.4.5 Limitations;84
7.2.2.5;6.5 Future Considerations;85
7.2.2.6;Acknowledgments;85
7.2.2.7;References;86
7.2.3;7. Preparation of Fluorescent Antibody Conjugate in Goats;88
7.2.3.1;7.1 Introduction;89
7.2.3.2;7.2 Materials;89
7.2.3.2.1;7.2.1 Reagents;89
7.2.3.2.2;7.2.2 Equipment;90
7.2.3.2.3;7.2.3 Biological Materials;90
7.2.3.3;7.3 Methods;90
7.2.3.3.1;7.3.1 Virus Isolation and Titration;90
7.2.3.3.1.1;7.3.1.1 General Principles and Remarks;90
7.2.3.3.1.2;7.3.1.2 Procedure;91
7.2.3.3.2;7.3.2 Virus Propagation and Cell Harvesting;92
7.2.3.3.3;7.3.3 Ribonucleoprotein Purification;93
7.2.3.3.4;7.3.4 Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Western Blotting;94
7.2.3.3.5;7.3.5 Immunization of Animals;96
7.2.3.4;7.4 Discussion;97
7.2.3.4.1;7.4.1 Experimental Tips;97
7.2.3.4.1.1;7.4.1.1 Viral Isolation and Titration;97
7.2.3.4.1.2;7.4.1.2 Ribonucleoprotein Purification;97
7.2.3.4.1.3;7.4.1.3 Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Western Blot Analysis;98
7.2.3.4.2;7.4.2 Critical Parameters and Troubleshooting;98
7.2.3.4.3;7.4.3 Precautions;98
7.2.3.4.4;7.4.4 Alternative Materials and Methods;98
7.2.3.4.5;7.4.5 Time Considerations;98
7.2.3.4.6;7.4.6 Limitations;98
7.2.3.5;Acknowledgments;98
7.2.3.6;References;100
7.2.4;8. Direct Fluorescent Antibody Test for Rabies Diagnosis;102
7.2.4.1;8.1 Introduction;102
7.2.4.2;8.2 Materials;104
7.2.4.2.1;8.2.1 Reagents;104
7.2.4.2.2;8.2.2 Supplies/Equipment;104
7.2.4.2.3;8.2.3 Biological Materials;105
7.2.4.3;8.3 Methods;105
7.2.4.4;8.4 Discussion;109
7.2.4.5;Acknowledgments;110
7.2.4.6;References;111
7.2.5;9. Use of a Rapid Skin Biopsy Technique for Human Rabies Antemortem Diagnosis;112
7.2.5.1;9.1 Introduction;112
7.2.5.2;9.2 Materials AND Methods;113
7.2.5.2.1;9.2.1 Equipment and Supplies;113
7.2.5.2.2;9.2.2 Protocol;117
7.2.5.2.2.1;9.2.2.1 Skin Biopsy;117
7.2.5.2.2.2;9.2.2.2 Tissue Preparation;118
7.2.5.2.2.3;9.2.2.3 Sectioning the Embedded Specimen;119
7.2.5.2.2.4;9.2.2.4 Acetone Fixation;121
7.2.5.2.2.5;9.2.2.5 Staining;121
7.2.5.2.2.6;9.2.2.6 Conjugate Titration;121
7.2.5.2.2.7;9.2.2.7 Microscopic Interpretation;122
7.2.5.3;9.3 Discussion;122
7.2.5.4;Acknowledgments;125
7.2.5.5;References;125
7.2.6;10. Immunohistochemistry;128
7.2.6.1;10.1 Introduction;128
7.2.6.2;10.2 Materials;129
7.2.6.2.1;10.2.1 Reagents;129
7.2.6.2.2;10.2.2 Equipment;129
7.2.6.2.3;10.2.3 Biological Materials;130
7.2.6.3;10.3 Methods;130
7.2.6.4;10.4 Discussion;132
7.2.6.4.1;10.4.1 Critical Parameters and Troubleshooting;132
7.2.6.4.2;10.4.2 Precautions;132
7.2.6.4.3;10.4.3 Alternative Materials and Methods;133
7.2.6.4.4;10.4.4 Time Considerations;133
7.2.6.4.5;10.4.5 Future Considerations;133
7.2.6.5;Acknowledgments;134
7.2.6.6;References;134
7.3;Section C Demonstration of Viral Nucleic Acids;136
7.3.1;11. Gel-Based Reverse Transcription-Polymerase Chain Reaction;138
7.3.1.1;11.1 Background;138
7.3.1.2;11.2 Preparatory Work and Procedures;140
7.3.1.2.1;11.2.1 Samples and Controls;140
7.3.1.2.2;11.2.2 RNA Extraction;140
7.3.1.2.3;11.2.3 Reverse Transcription of Viral RNA;140
7.3.1.2.4;11.2.4 Thermal Cycler;141
7.3.1.3;11.3 Reverse Transcription-Polymerase Chain Reaction Methodology;141
7.3.1.3.1;11.3.1 Amplification;142
7.3.1.3.2;11.3.2 Analyses of Reverse Transcription-Polymerase Chain Reaction Products by Electrophoresis on Agarose Gels;144
7.3.1.4;11.4 Discussion;145
7.3.1.5;Acknowledgments;146
7.3.1.6;References;146
7.3.2;12. Demonstration of Rabies Viral Nucleic Acids by In-Situ Polymerase Chain Reaction;148
7.3.2.1;12.1 Introduction;148
7.3.2.2;12.2 Materials;150
7.3.2.2.1;12.2.1 Reagents;150
7.3.2.2.2;12.2.2 Equipment;151
7.3.2.2.3;12.2.3 Biological Materials;151
7.3.2.3;12.3 Methods;151
7.3.2.3.1;12.3.1 Preparation of Samples (Impressions/Section/Infected Cells);151
7.3.2.3.2;12.3.2 Pre-Treatment with Proteinase K;152
7.3.2.3.3;12.3.3 Direct In-Situ Reverse Transcription and Amplification;153
7.3.2.3.4;12.3.4 Detection of Direct In-Situ Polymerase Chain Reaction-Amplified Targets;154
7.3.2.3.5;12.3.5 Interpretation of Results;155
7.3.2.4;12.4 Discussion;155
7.3.2.4.1;12.4.1 Experimental Tips;155
7.3.2.4.2;12.4.2 Critical Parameters and Troubleshooting;156
7.3.2.4.3;12.4.3 Precautions;157
7.3.2.4.4;12.4.4 Alternative Materials and/or Methods;157
7.3.2.4.5;12.4.5 Time Considerations;157
7.3.2.4.6;12.4.6 Limitations;158
7.3.2.4.7;12.4.7 Future Considerations;158
7.3.2.5;Acknowledgments;158
7.3.2.6;References;159
7.4;Section D Demonstration of Viral Antibodies: Binding Assays;162
7.4.1;13. Detection of Rabies Virus Antibodies by Competitive Enzyme-Linked Immunosorbent Assay;164
7.4.1.1;13.1 Introduction;164
7.4.1.2;13.2 Materials;166
7.4.1.2.1;13.2.1 Reagents and Plasticware;166
7.4.1.2.2;13.2.2 Equipment;166
7.4.1.2.3;13.2.3 Biological Materials;167
7.4.1.3;13.3 Methods;167
7.4.1.3.1;13.3.1 Antigen Preparation;167
7.4.1.3.2;13.3.2 Coating Plates with Antigen;167
7.4.1.3.3;13.3.3 Assay Procedure;168
7.4.1.3.4;13.3.4 Calculations, Test Acceptance, and Interpretation of Results;169
7.4.1.3.5;13.3.5 Assay Validation;170
7.4.1.4;13.4 Discussion;170
7.4.1.4.1;13.4.1 Experimental Tips;170
7.4.1.4.2;13.4.2 Critical Parameters and Troubleshooting;171
7.4.1.4.3;13.4.3 Precautions;172
7.4.1.4.4;13.4.4 Alternative Methods;172
7.4.1.4.5;13.4.5 Time Considerations;172
7.4.1.4.6;13.4.6 Limitations;172
7.4.1.5;Acknowledgments;172
7.4.1.6;References;174
7.4.2;14. An Indirect Fluorescent Antibody Test for the Serological Detection of Rabies Virus Immunoglobulin G and Immunoglo ...;176
7.4.2.1;14.1 Introduction;177
7.4.2.2;14.2 Materials;178
7.4.2.2.1;14.2.1 Equipment;178
7.4.2.2.2;14.2.2 Supplies;178
7.4.2.2.3;14.2.3 Reagents;179
7.4.2.2.4;14.2.4 Biological Materials;179
7.4.2.3;14.3 Methods;179
7.4.2.3.1;14.3.1 Specimen Collection and Handling;179
7.4.2.3.2;14.3.2 Preparation of Antigen Control Slides;180
7.4.2.3.3;14.3.3 Positive and Negative Control Samples;181
7.4.2.3.4;14.3.4 Indirect Fluorescent Antibody Standard Test Procedure;181
7.4.2.3.5;14.3.5 Reading Slides and Test Interpretation;182
7.4.2.3.6;14.3.6 Overall Interpretation;184
7.4.2.3.7;14.3.7 Procedures for Abnormal Results;186
7.4.2.4;14.4 Discussion;187
7.4.2.4.1;14.4.1 Considerations and Potential Limitations of the Indirect Fluorescent Antibody Test;187
7.4.2.5;14.5 Future Considerations;188
7.4.2.6;Acknowledgments;189
7.4.2.7;References;190
7.4.3;15. Demonstration of Viral Antibodies by the Counterimmunoelectrophoresis Test;194
7.4.3.1;15.1 Introduction;195
7.4.3.2;15.2 Materials;195
7.4.3.2.1;15.2.1 Reagents;195
7.4.3.2.2;15.2.2 Main Equipment;196
7.4.3.2.3;15.2.3 Biological Materials;196
7.4.3.2.4;15.2.4 Laboratory Animals;197
7.4.3.2.5;15.2.5 Specific Materials;197
7.4.3.3;15.3 Methods;197
7.4.3.3.1;15.3.1 Antigen Production;197
7.4.3.3.2;15.3.2 Production of the Hyper-Immune Indicator Serum;198
7.4.3.3.2.1;15.3.2.1 Preparation of the Vaccine;198
7.4.3.3.2.2;15.3.2.2 Immunization of Rabbits;198
7.4.3.3.2.3;15.3.2.3 Preparation of Agarose Gel Slides for Counterimmunoelectrophoresis Test and Titration of Antigen and Indicator Ser ...;198
7.4.3.3.2.4;15.3.2.4 Antigen Standardization;199
7.4.3.3.2.5;15.3.2.5 Hyper-Immune Indicator Serum Standardization;200
7.4.3.3.2.6;15.3.2.6 Counterimmunoelectrophoresis Technique;200
7.4.3.3.2.7;15.3.2.7 Reading and Interpreting the Results;200
7.4.3.4;15.4 Discussion;201
7.4.3.4.1;15.4.1 Experimental Tips;202
7.4.3.5;Acknowledgments;203
7.4.3.6;References;204
7.5;Section E Demonstration of Viral Antibodies: Functional Assays;206
7.5.1;16. The Mouse Neutralization Test;208
7.5.1.1;16.1 Introduction;208
7.5.1.2;16.2 Materials;209
7.5.1.2.1;16.2.1 Reagents;209
7.5.1.2.2;16.2.2 Equipment;209
7.5.1.2.3;16.2.3 Biological Materials;209
7.5.1.2.4;16.2.4 Laboratory Animals;210
7.5.1.2.5;16.2.5 Laboratory Supplies;210
7.5.1.3;16.3 Methods;210
7.5.1.3.1;16.3.1 Amplification of Rabies Virus;210
7.5.1.3.2;16.3.2 Titration of Rabies Virus;211
7.5.1.3.3;16.3.3 Neutralization of Rabies Virus by Antibodies;211
7.5.1.3.4;16.3.4 Interpretation of Results;212
7.5.1.4;16.4 Discussion;212
7.5.1.4.1;16.4.1 Experimental Tips;212
7.5.1.4.2;16.4.2 Critical Parameters and Troubleshooting;213
7.5.1.4.3;16.4.3 Precautions;214
7.5.1.4.4;16.4.4 Alternative Materials and/or Methods;214
7.5.1.4.5;16.4.5 Time Considerations;214
7.5.1.4.6;16.4.6 Limitations;215
7.5.1.5;References;216
7.5.2;17. The Rapid Fluorescent Focus Inhibition Test;218
7.5.2.1;17.1 Introduction;218
7.5.2.2;17.2 Materials;219
7.5.2.2.1;17.2.1 Reagents;219
7.5.2.2.2;17.2.2 Equipment;220
7.5.2.2.3;17.2.3 Biological Material;220
7.5.2.2.3.1;17.2.3.1 Cell Cultures;220
7.5.2.2.3.2;17.2.3.2 Rabies Virus;221
7.5.2.2.3.2.1;17.2.3.2.1 Titer Measurement of the Source Virus;221
7.5.2.2.3.2.2;17.2.3.2.2 Production of the Seed Virus;223
7.5.2.2.3.2.3;17.2.3.2.3 Production of the Stock Rabies Virus;223
7.5.2.2.3.2.4;17.2.3.2.4 Rapid Fluorescent Focus Inhibition Test Challenge Virus Dilution;223
7.5.2.2.3.3;17.2.3.3 Standard Rabies Immune Globulin;224
7.5.2.2.3.4;17.2.3.4 Fluorescein Isothiocyanate-Labeled Anti-Rabies Conjugate;224
7.5.2.2.4;17.2.4 Procedural Steps;224
7.5.2.2.4.1;17.2.4.1 Rapid Fluorescent Focus Inhibition Test Control Systems;224
7.5.2.2.4.2;17.2.4.2 Sample Preparation;225
7.5.2.2.4.3;17.2.4.3 Sample Dilution;225
7.5.2.2.4.4;17.2.4.4 Working Virus Dilution Preparation;226
7.5.2.2.4.5;17.2.4.5 Cell Suspension Preparation;226
7.5.2.2.4.6;17.2.4.6 Acetone Fixation of Slides and Staining;226
7.5.2.2.4.7;17.2.4.7 Microscopic Evaluation;227
7.5.2.2.4.8;17.2.4.8 Determination of Rabies Virus Neutralizing Antibody 50% Endpoint Titers;227
7.5.2.2.4.9;17.2.4.9 Test Interpretation;230
7.5.2.3;17.3 Discussion;231
7.5.2.4;Acknowledgments;232
7.5.2.5;References;233
7.5.3;18. The Fluorescent Antibody Virus Neutralization Test;236
7.5.3.1;18.1 Introduction;237
7.5.3.2;18.2 Materials;238
7.5.3.2.1;18.2.1 Reagents;238
7.5.3.2.2;18.2.2 Equipment;238
7.5.3.2.3;18.2.3 Biological Materials;239
7.5.3.3;18.3 Methods;239
7.5.3.3.1;18.3.1 Production of CVS-11 Virus on Cells;239
7.5.3.3.1.1;18.3.1.1 Growth of Cells;239
7.5.3.3.1.2;18.3.1.2 Infection of Cells;239
7.5.3.3.1.3;18.3.1.3 Virus Growth;239
7.5.3.3.1.4;18.3.1.4 Harvest and Storage;239
7.5.3.3.1.5;18.3.1.5 Determination of Virus Titer by TCID50 Assay (50% Tissue Culture Infective Dose);240
7.5.3.3.2;18.3.2 Fluorescent Antibody Virus Neutralization Test;240
7.5.3.3.2.1;18.3.2.1 Distribution of the Cell Culture Medium;240
7.5.3.3.2.1.1;18.3.2.1.1 For the Control Plate;241
7.5.3.3.2.1.2;18.3.2.1.2 For the Plates with Sera to be Tested;241
7.5.3.3.2.2;18.3.2.2 Distribution and Dilution of the Reference and Test Sera;241
7.5.3.3.2.3;18.3.2.3 Addition of Challenge Virus Standard;243
7.5.3.3.2.3.1;18.3.2.3.1 For the Plate with Sera to be Tested;243
7.5.3.3.2.3.2;18.3.2.3.2 For the Control Plate;243
7.5.3.3.2.4;18.3.2.4 Incubation of the Plates;243
7.5.3.3.2.5;18.3.2.5 Distribution of the Cell Suspension;244
7.5.3.3.2.6;18.3.2.6 Incubation of the Plates;244
7.5.3.3.2.7;18.3.2.7 Fixation;244
7.5.3.3.2.8;18.3.2.8 Staining;244
7.5.3.3.2.9;18.3.2.9 Reading;244
7.5.3.3.2.10;18.3.2.10 Calculation of the Titers;245
7.5.3.3.2.11;18.3.2.11 Conversion of the Titers;245
7.5.3.3.2.12;18.3.2.12 Validation of the Tests;246
7.5.3.4;18.4 Discussion;246
7.5.3.4.1;18.4.1 Critical Parameters;247
7.5.3.4.2;18.4.2 Precautions;247
7.5.3.4.3;18.4.3 Limitations;248
7.5.3.5;18.5 Future Considerations;248
7.5.3.6;Acknowledgements;248
7.5.3.7;References;248
7.6;Section F Typing/Differentiation of Lyssaviruses;252
7.6.1;19. Antigenic Typing of Lyssaviruses by Monoclonal Antibodies;254
7.6.1.1;19.1 Introduction;254
7.6.1.2;19.2 Materials;255
7.6.1.3;19.3 Methods;256
7.6.1.3.1;19.3.1 Preparation of the Viral Isolate for Analysis with Monoclonal Antibodies;256
7.6.1.3.1.1;19.3.1.1 Sample Preparation in Central Nervous System Tissue;256
7.6.1.3.1.2;19.3.1.2 Sample Preparation in Cell Culture;257
7.6.1.3.1.3;19.3.1.3 Titration of Monoclonal Antibodies;257
7.6.1.3.1.4;19.3.1.4 Titration of Conjugated Anti-Mouse Immune Globulin G;258
7.6.1.3.1.5;19.3.1.5 Application of the Technique;258
7.6.1.3.1.6;19.3.1.6 Interpretation of Results;259
7.6.1.4;19.4 Discussion;259
7.6.1.5;References;263
8;PART TWO Rabies Biologics;266
8.1;Section G Rabies Vaccines for Humans or Other Animals;268
8.1.1;20. Purified Chick-Embryo Cell Vaccine;270
8.1.1.1;20.1 Introduction;270
8.1.1.2;20.2 History;271
8.1.1.3;20.3 Preparation of the Vaccine;271
8.1.1.3.1;20.3.1 Seed Lot of Virus;271
8.1.1.3.2;20.3.2 Cell Cultures;272
8.1.1.3.3;20.3.3 Infection of Cells and Harvest of the Virus;272
8.1.1.3.4;20.3.4 Clarification of the Virus;273
8.1.1.3.5;20.3.5 Inactivation of the Virus;273
8.1.1.3.6;20.3.6 Concentration and Purification of the Virus;273
8.1.1.3.7;20.3.7 Preparation of the Final Vaccine;273
8.1.1.4;20.4 Control Tests;274
8.1.1.4.1;20.4.1 In-Process Controls;274
8.1.1.4.2;20.4.2 Potency Tests;274
8.1.1.4.3;20.4.3 Stability Test;274
8.1.1.4.4;20.4.4 Tests on the Cell Culture;274
8.1.1.4.5;20.4.5 Identity Testing;274
8.1.1.5;20.5 Administration of the Vaccine;275
8.1.1.5.1;20.5.1 Special Precautions for the Intradermal Route of Administration;275
8.1.1.5.2;20.5.2 Pre-Exposure Prophylaxis;275
8.1.1.5.3;20.5.3 Post-Exposure Prophylaxis;275
8.1.1.6;20.6 Presentation and Storage;276
8.1.1.7;20.7 Laboratory Tests;276
8.1.1.8;References;278
8.1.2;21. Purified Vero-Cell Rabies Vaccine;280
8.1.2.1;21.1 Introduction;280
8.1.2.2;21.2 Materials;281
8.1.2.2.1;21.2.1 Reagents and Culture Medium;281
8.1.2.2.2;21.2.2 Equipment;281
8.1.2.2.3;21.2.3 Biological Materials;282
8.1.2.2.3.1;21.2.3.1 Cell Seed;282
8.1.2.2.3.2;21.2.3.2 Virus Strain;282
8.1.2.2.3.3;21.2.3.3 Animals for Control Tests;282
8.1.2.2.3.4;21.2.3.4 Cell Lines for Control Tests;282
8.1.2.3;21.3 Methods;282
8.1.2.3.1;21.3.1 Preparation of Cell Banks;282
8.1.2.3.2;21.3.2 Preparation of the Virus Seed Banks;283
8.1.2.3.3;21.3.3 Cell Culture System and Virus Production;283
8.1.2.3.4;21.3.4 Concentration and Purification of Harvested Virus Suspensions;284
8.1.2.3.5;21.3.5 Rabies Virus Inactivation;284
8.1.2.3.6;21.3.6 Preparation of the Final Bulk;284
8.1.2.3.7;21.3.7 Formulation and Filling of the Bulk Vaccine;284
8.1.2.3.8;21.3.8 Expiry Date;285
8.1.2.3.9;21.3.9 Quality Control and Quality Assurance;285
8.1.2.4;21.4 Discussion;285
8.1.2.5;Acknowledgments;286
8.1.2.6;References;286
8.1.3;22. Mouse Potency Testing of Rabies Vaccines;288
8.1.3.1;22.1 Introduction;288
8.1.3.2;22.2 Materials;289
8.1.3.2.1;22.2.1 Reagents;289
8.1.3.2.2;22.2.2 Equipment;290
8.1.3.2.3;22.2.3 Animals;290
8.1.3.2.4;22.2.4 Challenge Virus Standard;290
8.1.3.2.5;22.2.5 Reference Vaccine;290
8.1.3.2.6;22.2.6 Vaccines;291
8.1.3.3;22.3 Methods;291
8.1.3.3.1;22.3.1 Vaccination of Mice;291
8.1.3.3.2;22.3.2 Challenge;291
8.1.3.3.3;22.3.3 Monitoring;292
8.1.3.3.4;22.3.4 Euthanasia;293
8.1.3.3.5;22.3.5 Potency Determination;293
8.1.3.3.6;22.3.6 Validation of Potency Tests;293
8.1.3.3.7;22.3.7 Minimum Potency Requirements;294
8.1.3.4;22.4 Discussion;294
8.1.3.4.1;22.4.1 Critical Parameters;295
8.1.3.4.2;22.4.2 Precautions;295
8.1.3.4.3;22.4.3 Future Consideration;295
8.1.3.5;Acknowledgments;296
8.1.3.6;References;297
8.1.4;23. In Vitro Potency Testing of Rabies Vaccines;300
8.1.4.1;23.1 Introduction;300
8.1.4.2;23.2 Materials;301
8.1.4.2.1;23.2.1 Reagents;301
8.1.4.2.2;23.2.2 Equipment;302
8.1.4.2.3;23.2.3 Biological Materials;302
8.1.4.2.4;23.2.4 Laboratory Supplies;302
8.1.4.3;23.3 Methods;303
8.1.4.3.1;23.3.1 Coating of Enzyme-Linked Immunosorbent Assay Plates;303
8.1.4.3.2;23.3.2 Blocking;303
8.1.4.3.3;23.3.3 Preparation and Addition of Antigen Dilutions;303
8.1.4.3.4;23.3.4 Preparation and Addition of Detector Reagents;303
8.1.4.3.5;23.3.5 Interpretation of Results;304
8.1.4.3.5.1;23.3.5.1 Non-Linear Regression Curve Fitting Model;304
8.1.4.3.5.2;23.3.5.2 Four Parameter Logistic Equation;305
8.1.4.3.5.3;23.3.5.3 Five Parameter Logistic Equation;305
8.1.4.3.5.4;23.3.5.4 Parallel Line Bioassay Method;305
8.1.4.4;23.4 Discussion;306
8.1.4.4.1;23.4.1 Experimental Tips and Precautions;306
8.1.4.4.2;23.4.2 Critical Parameters and Troubleshooting;306
8.1.4.4.3;23.4.3 Alternative Materials and/or Methods;306
8.1.4.4.4;23.4.4 Time Considerations;306
8.1.4.4.5;23.4.5 Limitations;307
8.1.4.5;References;307
8.2;Section H Rabies Antibodies/Fragments;310
8.2.1;24. Production of Equine Rabies Immune Globulin of High Purity, Potency, and Safety;312
8.2.1.1;24.1 Introduction;313
8.2.1.2;24.2 Materials;313
8.2.1.2.1;24.2.1 Biological Materials;313
8.2.1.2.2;24.2.2 Reagents;313
8.2.1.2.3;24.2.3 Laboratory Animals;313
8.2.1.2.4;24.2.4 Equipment for Production;314
8.2.1.2.5;24.2.5 Equipment for Testing;314
8.2.1.3;24.3 Methods;314
8.2.1.3.1;24.3.1 Production Method;314
8.2.1.3.2;24.3.2 Process Validation of Production Batches;316
8.2.1.3.3;24.3.3 Study Design;316
8.2.1.3.4;24.3.4 Potency Assay by the Rapid Fluorescence Focus Inhibition Test;316
8.2.1.3.5;24.3.5 Content of F(ab’)2 and Impurities;317
8.2.1.3.6;24.3.6 Safety, Pharmacokinetics, Immunogenicity, and Animal Efficacy of the Thai Red Cross Society Equine Rabies Immune Glo ...;317
8.2.1.3.6.1;24.3.6.1 Pharmacokinetic Analysis;317
8.2.1.3.6.2;24.3.6.2 Purity Study of the Thai Red Cross Society Equine Rabies Immune Globulin;318
8.2.1.3.6.2.1;24.3.6.2.1 Determination of the Content of F(ab’)2 and Impurities;318
8.2.1.3.6.2.2;24.3.6.2.2 Accelerated Stability Study of the Thai Red Cross Society Equine Rabies Immune Globulin;320
8.2.1.3.6.2.3;24.3.6.2.3 Long-Term Stability Results of the Thai Red Cross Society Equine Rabies Immune Globulin (Real-Time);320
8.2.1.4;24.4 Discussion;321
8.2.1.5;Acknowledgments;321
8.2.1.6;References;322
8.2.2;25. Production of Polyclonal Rabies Virus Antibodies in Birds;324
8.2.2.1;25.1 Introduction;324
8.2.2.2;25.2 Advantages of Immunoglobulin Y Compared to Immunoglobulin G;325
8.2.2.3;25.3 Materials and Methods;327
8.2.2.3.1;25.3.1 Immunization of Rabies Virus Antigens;327
8.2.2.3.2;25.3.2 Preparation of Rabies Virus Antigens;327
8.2.2.3.3;25.3.3 Preparation of Water in Oil Emulsion of Antigens;328
8.2.2.3.4;25.3.4 Immunization of Hens and Collection of Samples;328
8.2.2.3.5;25.3.5 Determination of Antibody Titers in Egg Yolk;328
8.2.2.3.6;25.3.6 Purification of Immunoglobulin Y;329
8.2.2.3.7;25.3.7 Egg Production After Immunization;330
8.2.2.3.8;25.3.8 Changes of Antibody Titers;330
8.2.2.3.9;25.3.9 Immunoglobulin Y Purification and its High-Performance Liquid Chromatography Profile;330
8.2.2.4;25.4 Discussion;333
8.2.2.5;References;334
8.2.3;26. Production of Full-Length Human Monoclonal Antibodies using Transgenic Mice;336
8.2.3.1;26.1 Introduction;336
8.2.3.2;26.2 Hybridoma Production;339
8.2.3.2.1;26.2.1 Mouse Immunization;339
8.2.3.2.2;26.2.2 Screening for Antibody Responses in Humanized Mice;340
8.2.3.2.3;26.2.3 Hybridoma Fusions;340
8.2.3.3;26.3 Hybridoma Screening and Characterization;342
8.2.3.3.1;26.3.1 Cell Culture and Screening;342
8.2.3.3.2;26.3.2 Virus Neutralization;343
8.2.3.3.3;26.3.3 Pseudovirus Neutralization;343
8.2.3.3.4;26.3.4 Epitope Mapping;344
8.2.3.3.5;26.3.5 Further Characterization;344
8.2.3.3.6;26.3.6 Hamster Post-Exposure Prophylaxis;345
8.2.3.4;26.4 Summary;345
8.2.3.5;References;346
9;PART THREE Development of Anti-Viral Approaches;348
9.1;27. Rabies Therapeutics: Development of Anti-Viral Approaches;350
9.1.1;27.1 Background;351
9.1.2;27.2 An Empirical and Heteroclite Arsenal;352
9.1.3;27.3 Blocking External Replication Steps;355
9.1.3.1;27.3.1 Antibodies;355
9.1.3.2;27.3.2 Dermaseptins;356
9.1.3.3;27.3.3 Aptamers;357
9.1.4;27.4 Blocking Internal Replication Steps;357
9.1.4.1;27.4.1 Viral Transport;357
9.1.4.2;27.4.2 Viral Transcription/Replication;357
9.1.4.2.1;27.4.2.1 Inhibiting Identified Functional Interaction;358
9.1.4.2.2;27.4.2.2 RNA Interference and Antisense;359
9.1.5;27.5 Screening Without “A Priori” Knowledge;359
9.1.5.1;27.5.1 Yeast 2-Hybrid to Screen Libraries of Peptides Able to Destabilize the Protein–Protein Interactions;360
9.1.5.2;27.5.2 Phage Display to Screen Libraries of Small Peptides Able to Interact with the N-RNA Template of Rabies Virus;361
9.1.5.3;27.5.3 High-Throughput Screening in Infected Cells or Cell-Free Systems;361
9.1.6;27.6 Several Methodologies;362
9.1.6.1;27.6.1 Rabies Virus Minireplicon System;362
9.1.6.2;27.6.2 Phage Display Screening;363
9.1.6.2.1;27.6.2.1 Library Construction;363
9.1.6.2.2;27.6.2.2 Selection Process;364
9.1.6.2.3;27.6.2.3 Monoclonal Phage Production and Measurement of the Affinity by Enzyme-Linked Immunosorbent Assay;365
9.1.7;27.7 Discussion;365
9.1.8;Acknowledgments;366
9.1.9;References;367
10;Appendix A: List of Biologics Recommended by WHO for Human Rabies Prophylaxis;372
11;Index;374

Leseproben

List of Contributors
Surasak Akesowan,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand C. Balachandran,     Department of Veterinary Pathology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai, India Mohamed Ben Mechlia,     Antiviral Strategies Unit, Institut Pasteur, Paris Cedex 15, France Hervé Bourhy,     Institut Pasteur, Unit Lyssavirus Dynamics and Host Adaptation, National Reference Centre for Rabies, WHO Collaborative Center for Reference and Research on Rabies, Paris Cedex 15, France Graciane Caporale,     Instituto Pasteur, São Paulo, São Paulo, Brazil Guillaume Castel,     INRA, UMR 1062 CBGP, Montferrier-sur-Lez, France Florence Cliquet,     French Agency for Food, Environmental and Occupational Health and Safety, Nancy Laboratory for Rabies and Wildlife, European Union Reference Laboratory for Rabies, European Union Reference Laboratory for Rabies Serology, OIE Reference Laboratory for Rabies, Technopôle Agricole et Vétérinaire, Malzéville, France Laurent Dacheux,     Institut Pasteur, Unit Lyssavirus Dynamics and Host Adaptation, National Reference Centre for Rabies, WHO Collaborative Center for Reference and Research on Rabies, Paris Cedex 15, France Svastijaya Daviratanasilpa,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand Bernhard Dietzschold,     Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA Clifton P. Drew,     Infectious Diseases Pathology Branch, Centers for Disease Control and Prevention, Atlanta, GA, USA Christine Fehlner-Gardiner,     Canadian Food Inspection Agency, Centre of Expertise for Rabies, Ottawa, Canada Anthony R. Fooks Wildlife Zoonosis and Vector-Borne Diseases Research Group, Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, UK Institute of Infection and Global Health, University of Liverpool, Liverpool, UK Neuza Maria Frazatti-Gallina,     Instituto Butantan, São Paulo, Brazil Conrad M. Freuling,     Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Molecular Virology and Cell Biology, Greifswald-Insel Riems, Germany Bin Gao,     The Centre for Molecular Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China Cynthia S. Goldsmith,     Infectious Diseases Pathology Branch, Centers for Disease Control and Prevention, Atlanta, GA, USA Hajime Hatta,     Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan Wachiraporn Hemmala,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand Satoshi Inoue,     Department of Food and Nutrition, Faculty of Home Economics, Kyoto Women’s University, Kyoto, Japan Corinne Jallet,     Antiviral Strategies Unit, Institut Pasteur, Paris Cedex 15, France Pakamatz Khawplod,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand Sumana Khomvilai,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand Ivanete Kotait,     Instituto Pasteur, São Paulo, Brazil Ivan V. Kuzmin,     Global Alliance for Rabies Control, Manhattan, KS, USA Kornvika Limsuwun,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand Maria Luiza Carrieri,     Rabies Laboratory, Butantan Institute, São Paulo, São Paulo, Brazil Claudius Malerczyk,     Novartis Vaccines and Diagnostics GmbH, Marburg, Germany Bonny Mayes,     Texas Department of State Health Services, Texas, USA Lorraine M. McElhinney,     Wildlife Zoonosis and Vector-Borne Diseases Research Group, Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, UK Sharon Messenger,     Viral and Rickettsial Disease Laboratory, California Department of Public Health, Richmond, CA, USA Susan M. Moore,     Kansas State University Rabies Laboratory, Manhattan, KS, USA Wildeberg C. Moreira,     Fundação Oswaldo Cruz, Rio de Janeiro, Brazil Wlamir C. Moura,     Fundação Oswaldo Cruz, Rio de Janeiro, Brazil Helmut Müller,     Novartis Vaccines and Diagnostics GmbH, Marburg, Germany Thomas F. Müller,     Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Molecular Virology and Cell Biology, Greifswald-Insel Riems, Germany Thirumeni Nagarajan,     Biological E. Limited, Shameerpet, Hyderabad, India Ernest Ngoepe,     OIE Rabies Reference Laboratory, ARC-Onderstepoort Veterinary Institute Onderstepoort, South Africa Naruemol Pakmanee,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand Chun-Ho Park,     Department of Veterinary Pathology, School of Veterinary Medicine, Kitasato University, Towada, Japan Zélia Peixoto,     Instituto Pasteur, São Paulo, São Paulo, Brazil Duangporn Pornmuttakun,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand Mingsheng Qu,     Anhui Medical University, Anhui, China Luzia H. Queiroz,     University Est. Paulista, Faculdade de Medicina Veterinária de Araçatuba, Departamento de Apoio, Produção e Saúde Animal, Araçatuba, São Paulo, Brazil G. Dhinakar Raj,     Translational Research Platform for Veterinary Biologicals, Tamil Nadu Veterinary and Animal Sciences University, Madhavaram Milk Colony, Chennai, India Angannan Rajasekaran,     Biological E Limited, Hyderabad, India Rajendran Ramya,     Indian Immunologicals Limited, Gachibowli, Hyderabad, India Robert J. Rudd,     New York State Department of Health, Albany, New York, USA Charles E. Rupprecht,     Global Alliance for Rabies Control, Manhattan, KS, USA, Ross University School of Veterinary Medicine, Basseterre, St. Kitts, West Indies Claude Sabeta,     OIE Rabies Reference Laboratory, ARC-Onderstepoort Veterinary Institute Onderstepoort, South Africa Lalida Sakolpap,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand Alexandre Servat,     French Agency for Food, Environmental and Occupational Health and Safety, Nancy Laboratory for Rabies and Wildlife, European Union Reference Laboratory for Rabies, European Union Reference Laboratory for Rabies Serology, OIE Reference Laboratory for Rabies, Technopôle Agricole et Vétérinaire, Malzéville, France Wun-Ju Shieh,     Infectious Diseases Pathology Branch, Centers for Disease Control and...