Dengue, 2014
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1138

Infection by flaviviruses such as dengue virus serotypes (DENV 1-4), Japanese encephalitis virus (JEV), tick-borne encephalitis virus (TBE), yellow fever virus (YFV) and West Nile virus (WNV) impact millions of lives and cause tens of thousands of mortalities each year. Dengue is a global public health emergency especially since there is no preventative vaccine or antiviral treatment for dengue disease. Dengue: Methods and Protocols offers the increasing number of dengue researchers a one-stop protocol book with techniques compiled from the leading laboratories working on dengue. Chapters cover topics such as dengue virus isolation from clinical samples, quantification of human antibodies against the virus, assays to quantify the virus particles, the widely used mouse model to study dengue pathogenesis, vaccine and antiviral efficacies. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Dengue: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies on dengue research.

Dengue Virus Growth, Purification and Fluorescent Labeling.- Isolation and Titration of Dengue Viruses by the Mosquito Inoculation Technique.- Measuring Antibody Neutralization of Dengue Virus (DENV) Using a Flow Cytometry-Based Technique.- Dengue Virus Purification and Sample Preparation for Cryo-Electron Microscopy.- Development of a Multiplex Bead-Based Assay to Monitor Dengue Virus Seroconversion.- Pseudo-Infectious Reporter Virus Particles for Measuring Antibody-Mediated

Neutralization and Enhancement of Dengue Virus Infection.- Cell-Based Flavivirus Infection (CFI) Assay for the Evaluation of Dengue Antiviral Candidates Using High Content Imaging.- Development and Application of Dengue Virus Reverse Genetic Systems.- DENV4 Renilla Luciferase Reporter Replicons.- Targeted Mutagenesis of Dengue Virus Type 2 replicon RNA by Yeast in vivo

Recombination.- Identification of Dengue-Specific Human Antibody Fragments Using Phage Display.- Next Generation Whole Genome Sequencing of Dengue Virus.- MPGAfold in Dengue Secondary Structure Prediction.- Insights into Secondary and Tertiary Interactions of Dengue Virus RNA by SHAPE.- Use of Small Angle X-Ray Scattering to Investigate the Structure and Function of Dengue Virus NS3 and NS5.- Identification of Dengue RNA Binding Proteins Using RNA Chromatography and Quantitative Mass Spectrometry.- Analysis of Affinity of Dengue Virus Protein Interaction Using BIAcore.- Functional Genomics Approach for the Identification of Human Host Factors Supporting Dengue Viral Propagation.- Investigating Dengue Virus Non-Structural Protein 5 (NS5) Nuclear Import.- Small Molecule Inhibitor Discovery for Dengue Virus Protease Using High Throughput Screening.- Construction of Dengue Virus Protease Expression Plasmid and in vitro Protease Assay for Screening Antiviral Inhibitors.- Construction of Plasmid, Bacterial Expression, Purification, and Assay of Dengue Virus Type 2 NS5 Methyltransferase.- Animal Models in Dengue.- Evaluation of Dengue Antiviral Candidates In Vivo in Mouse Model.