Main description:

This volume provides various techniques and methodologies currently used in the study of MERS-CoV. Chapters are divided into four parts detailing evolution and entry of MERS-coronavirus, genetic alteration and structural determination of MERS-coronavirus proteins, quantitation of virus and anti-viral factors, and mouse models for MERS -coronavirus. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, MERS Coronavirus: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

PART I: Evolution and Entry of MERS-Coronavirus

1. Studying Evolutionary Adaptation of MERS-CoV

Michael Letko and Vincent Munster

2. Evaluating MERS-CoV Entry Pathways

Enya Qing, Michael P Hantak, Gautami G Galpalli, and Tom Gallagher

3. Biochemical Characterization of Middle East Respiratory Syndrome Coronavirus Spike Protein Proteolytic Processing

Gary R. Whittaker and Jean K. Millet

4. Crystallization and Structural Determination of the Receptor-binding Domain of MERS-CoV Spike Glycoprotein

Haixia Zhou, Shuyuan Zhang, and Xinquan Wang

PART II: Genetic Alteration and Structural Determination of MERS-Coronavirus Proteins

5. Bacterial Artificial Chromosome-based Lambda Red Recombination with the I-SceI Homing Endonuclease for Genetic Alteration of MERS-CoV

Anthony R. Fehr

6. Deducing the Crystal Structure of MERS-CoV Helicase

Sheng Cui and Wei Hao

PART III: Quantitation of Virus and Anti-Viral Factors

7. Antigen Capture Enzyme-linked Immunosorbent Assay for Detecting Middle East Respiratory Syndrome Coronavirus in Humans

Joshua Fung, Susanna K. P. Lau, and Patrick C. Y. Woo

8. Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR

Abdullah Algaissi, Anurodh S. Agrawal, Anwar M. Hashem, and Chien-Te K. Tseng

9. Evaluation of MERS-CoV Neutralizing Antibodies in Sera Using Live Virus Microneutralization Assay Abdullah Algaissi and Anwar M Hashem

10. Generation of MERS-CoV Pseudotyped Viral Particles for the Evaluation of Neutralizing Antibodies in Mammalian Sera Abdulrahman Almasaud, Naif Khalaf Alharbi, and Anwar M Hashem

11. Qualitative and Quantitative Determination of MERS-CoV S1-specific Antibodies using ELISA

Sawsan S Al-amri and Anwar M Hashem

PART IV: Mouse Models for MERS -Coronavirus

12. Genetically Engineering a Susceptible Mouse Model for MERS-CoV-Induced

Acute Respiratory Distress Syndrome

Sarah R. Leist and Adam S. Cockrell

13. Development of a Mouse Adapted MERS Coronavirus

Kun Li and Paul B. McCray, Jr.

14. Metabolite, Protein, and Lipid Extraction (MPLEx): A Method that Simultaneously Inactivates Middle East Respiratory Syndrome Coronavirus and Allows Analysis of Multiple Host Cell Components Following Infection

Carrie D. Nicora, Amy C. Sims, Kent J. Bloodsworth, Young-Mo Kim, Ronald J. Moore, Jennifer E. Kyle, Ernesto S. Nakayasu, and Thomas O. Metz

15. Evaluation of Activation and Inflammatory Activity of Myeloid Cells During

Pathogenic Human Coronavirus Infection

Rudragouda Channappanavar and Stanley Perlman

16. Histopathologic Evaluation and Scoring of Viral Lung Infection David K. Meyerholz and Amanda P. Beck