Main description:

This detailed new edition provides a comprehensive collection of protocols applicable to all members of the Coronavirinae sub-family currently and that are also transferrable to other fields of virology. Beginning with a section on detection, discovery, and evolution, the volume continues with coverage of propagation and titration of coronaviruses, genome manipulation, study of virus-host interactions, as well as imaging coronavirus infections. 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 cutting-edge, Coronaviruses: Methods and Protocols, Second Edition serves as a valuable guide to researchers working to identify and control viruses with increased potential to cross the species barrier and to develop the diagnostics, vaccines, and antiviral therapeutics that are required to manage future outbreaks in both humans and animals.

Contents:

1. Coronaviruses: An Updated Overview of Their Replication and Pathogenesis

Yuhang Wang, Matthew Grunewald, and Stanley Perlman

Part I: Coronavirus Detection, Discovery, and Evolution

2. Discovery of Novel Coronaviruses in Rodents

Theocharis Tsoleridis and Jonathan K. Ball

3. Detection and Discovery of Coronaviruses in Wild Bird Populations

Chantal J. Snoeck and Siamak Zohari

4. Competitive ELISA for the Detection of Serum Antibodies Specific for Middle East Respiratory Syndrome-Coronavirus (MERS-CoV)

Shuetsu Fukushi

5. Whole Genome Sequencing Protocols for IBV and Other Coronaviruses Using High-Throughput Sequencing

Graham L. Freimanis and Michael S. Oade

Part II: Propagation and Titration of Coronaviruses

6. Isolation and Tissue Culture Adaptation of Porcine Deltacoronavirus: A Case Study

Hui Hu, Kwonil Jung, Scott P. Kenney, and Linda J. Saif

7. The Preparation of Chicken Kidney Cell Cultures for Virus Propagation

Elena Lokhman, Srijana Rai, and Billy Matthews

8. The Preparation of Chicken Tracheal Organ Cultures and Their Application for Ciliostasis Test, Growth Kinetics Studies, and Virus Propagation

Giulia Dowgier and Erica Bickerton

9. Isolation and Propagation of Coronaviruses in Embryonated Eggs

James S. Guy

10. Well-Differentiated Primary Mammalian Airway Epithelial Cell Cultures

Mitra Gultom, Laura Laloli, and Ronald Dijkman

11. Quantification of Coronaviruses by Titration In Vitro and Ex Vivo

Giulia Dowgier and Helena J. Maier

Part III: Manipulating the Genomes of Coronaviruses

12. Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses

Sarah Keep, Paul Britton, and Erica Bickerton

13. In-Yeast Assembly of Coronavirus Infectious cDNA Clones Using a Synthetic Genomics Pipeline

Tran Thi Nhu Thao, Fabien Labroussaa, Nadine Ebert, Joerg Jores, and Volker Thiel

Part IV: Studying Virus-Host Interactions

14. Proximity Labeling for Identification of Coronavirus-Host Protein Interactions

Philip V'kovski, Silvio Steiner, and Volker Thiel

15. Using Yeast to Identify Coronavirus-Host Protein Interactions

Stuart Weston and Matthew Frieman

16. Determining How Coronaviruses Overcome the Interferon and Innate Immune Response

Christopher M. Coleman

17. Ribopuromycylation in Coronavirus-Infected Cells

Matthew J. Brownsword, Helena J. Maier, and Nicolas Locker

Part V: Imaging Coronavirus Infections

18. Visualizing Coronavirus Entry into Cells

Aleksandra Milewska, Katarzyna Owczarek, Artur Szczepanski, and Krzysztof Pyrc

19. Preparation of Cultured Cells Using High Pressure Freezing and Freeze Substitution for Subsequent 2D or 3D Visualization in the Transmission Electron Microscope

Nicole Doyle and Philippa C. Hawes