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This detailed volume explores various aspects of the dengue virus and its four serotypes (DENV1-4). Beginning with a section on protocols to produce DENV and its proteins that are fundamental for many biophysical, biochemical, immunological, and cellular studies, the book continues by covering protocols to study the interactions between DENV and cellular proteins, DENV immunopathogenesis and diagnosis, as well as recent advances in animal models that can be used in studies of DENV immunopathogenesis and vaccine development. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Dengue Virus: Methods and Protocols serves as an ideal guide for researchers attempting to further understand this critical threat to worldwide public health.
List of contents
Virus Propagation and Plaque Assay for Dengue Virus.- Transmission Electron Microscopy Studies of Dengue Viruses.- Production of Properly Folded NS1 Protein in Bacterial Cells.- Expression and Purification of Dengue NS1 Protein in Sf9-Baculovirus System.- Purification of Dengue and Zika Virus Non-Structural Protein 5 for Crystallization and Screening of Antivirals.- Obtention of Dengue Virus Membrane Proteins and Role for Virus Assembly.- Construction of Genomic and Sub-Genomic Dengue Infectious Replicons.- Dengue Virus Capsid-Protein Dynamics in Live Infected Cells Studied by Pair Correlation Analysis.- Yeast Two-Hybrid System for Mapping Novel Dengue Protein Interactions.- Isolation and Identification of Dengue Virus Interactome with Human Plasma Proteins by Affinity Purification-Mass Spectrometry.- Molecular Diagnosis Of Dengue.- Serological Diagnosis of Dengue.- Measuring Transendothelial Electrical Resistance (TEER) for Dengue Infection Studies.- Evaluation of DENV-InducedEndothelial Cell Permeability by Measurements of Transendothelial Electrical Resistance (TEER) and Extravasation of Proteins and Virus.- In Vitro Cytokine Production by Dengue-Infected Human Monocyte-Derived Dendritic Cells.- In Vitro Generation of Human Antibody-Secreting Cells through the Stimulation of PBMCs with Dengue Virus Particles.- Isolation of Microvesicles from Plasma Samples Avoiding Lipoprotein Contamination.- Evaluating Dengue Virus Pathogenesis in Mice and Humans by Histological and Immunohistochemistry Approaches.- Humanized Mice for the Study of Dengue Disease Pathogenesis: Biological Assays.
