Coronaviruses: Methods and Protocols

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

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

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