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Medicine - Internal Medicine | Dengue - Methods and Protocols


Methods and Protocols

Series: Methods in Molecular Biology, Vol. 1138

Padmanabhan, Radhakrishnan, Vasudevan, Subhash G. (Eds.)

2014, XIII, 403 p. 82 illus., 49 illus. in color.

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  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

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.

Content Level » Professional/practitioner

Keywords » RNA proteins - antiviral screening - dengue virus - enzyme assays - virology

Related subjects » Biomedical Sciences - Internal Medicine - Virology

Table of contents 

Part I. Modern Quantitative Virological Methods


1. Dengue Virus Growth, Purification and Fluorescent Labeling

            Summer Zhang, Kuan Rong Chan, Hwee Cheng Tan, and Eng Eong Ooi


2. Isolation and Titration of Dengue Viruses by the Mosquito Inoculation Technique

            Milly M. Choy and Duane J. Gubler


3. Measuring Antibody Neutralization of Dengue Virus (DENV) Using a Flow Cytometry-Based Technique

            A. Ruklanthi de Alwis and Aravinda M. de Silva


4. Dengue Virus Purification and Sample Preparation for Cryo-Electron Microscopy

            Joanne L. Tan and Lok Shee Mei


5. Development of a Multiplex Bead-Based Assay to Monitor Dengue Virus Seroconversion

            Kaiting Ng and John E. Connolly


6. Pseudo-Infectious Reporter Virus Particles for Measuring Antibody-Mediated

Neutralization and Enhancement of Dengue Virus Infection

            Swati Mukherjee, Ted Pierson and Kimberly A. Dowd


7. Cell-Based Flavivirus Infection (CFI) Assay for the Evaluation of Dengue Antiviral Candidates Using High Content Imaging

            Kah Hin Tan, Kitti Chan Wing Ki, Satoru Watanabe, Subhash G. Vasudevan and Manoj Krishnan



Part II. Reverse Genetic Systems to Study Virus Replication and Evolution


8. Development and Application of Dengue Virus Reverse Genetic Systems

            Andrew Davidson


9. DENV4 Renilla Luciferase Reporter Replicons

            Sofia Alcaraz-Estrada, Rosa Del Angel, and R. Padmanabhan


10. Targeted Mutagenesis of Dengue Virus Type 2 replicon RNA by Yeast in vivo


            Mark Manzano and R. Padmanabhan


11. Identification of Dengue-Specific Human Antibody Fragments Using Phage Display

Moon Y.F. Tay, Chin Chin Lee, Subhash G. Vasudevan and Nicole J. Moreland


12. Next Generation Whole Genome Sequencing of Dengue Virus

Pauline Poh Kim Aw, Paola Florez de Sessions, Andreas Wilm, Long Truong Hoang, Niranjan Nagarajan, October M. Sessions, and Martin Lloyd Hibberd



Part III. Modern Methods to Study Conformation of RNA Proteins and Their Molecular Interactions


13. MPGAfold in Dengue Secondary Structure Prediction

            Wojciech K. Kasprzak and Bruce A. Shapiro


14. Insights into Secondary and Tertiary Interactions of Dengue Virus RNA by SHAPE

            Joanna Sztuba-Solinska and Stuart F.J. Le Grice


15. Use of Small Angle X-Ray Scattering to Investigate the Structure and Function of Dengue Virus NS3 and NS5

            Kyung H. Choi and Marc Morais


16. Identification of Dengue RNA Binding Proteins Using RNA Chromatography and Quantitative Mass Spectrometry

            A.M. Ward, J. Gunaratne, M. A. Garcia-Blanco


17. Analysis of Affinity of Dengue Virus Protein Interaction Using BIAcore

            Yau Yin Hoe and Susana Geifman Shochat


18. Functional Genomics Approach for the Identification of Human Host Factors Supporting Dengue Viral Propagation

            Nicholas J. Barrows, Sharon F. Jamison, Shelton S. Bradrick, Caroline Le Sommer, So Young Kim, James Pearson, and Mariano A. Garcia-Blanco


19. Investigating Dengue Virus Non-Structural Protein 5 (NS5) Nuclear Import

            Johanna E. Fraser, Stephen M. Rawlinson, Chunxiao Wang, David A. Jans, and Kylie M. Wagstaff



Part IV. In vitro Enzyme Assays for Antiviral Screening


20. Small Molecule Inhibitor Discovery for Dengue Virus Protease Using High Throughput Screening

            Mark Manzano, Janak Padia, and  R. Padmanabhan


21. Construction of Dengue Virus Protease Expression Plasmid and in vitro Protease Assay for Screening Antiviral Inhibitors

            Huiguo Lai, Tadahisa Teramoto and Radhakrishnan Padmanabhan


22. Construction of Plasmid, Bacterial Expression, Purification, and Assay of Dengue Virus Type 2 NS5 Methyltransferase

            Siwaporn Boonyasuppayakorn and Radhakrishnan Padmanabhan



Part V. Dengue Animal Model for Vaccines and Antivirals


23. Animal Models in Dengue

            Emily Plummer and Sujan Shresta


24. Evaluation of Dengue Antiviral Candidates In Vivo in Mouse Model

            Satoru Watanabe and Subhash G. Vasudevan

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