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This new edition explores and provides an update on the biology and pathogenesis of human cytomegalovirus infection.  Modern techniques that are currently being utilized to investigate the molecular aspects of viral infection, as well as how these new research studies are leading to new approaches to mitigate disease, are also provided. Given the key role the virus plays in significant acute and chronic human disease in all stages of life, from newborns to seniors, the need for clear methodologies to further explore the biology of HCMV infection and mitigation strategies is readily apparent. Written in the highly successful Methods in Molecular Biology 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 up-to-date, Human Cytomegaloviruses: Methods and Protocols, Second Edition serves as an ideal scientific reference for basic and clinical scientists and medical personnel on the modern understanding of the pathobiology of the virus, and the approaches, techniques, and models to study human cytomegalovirus infection and disease.

Human Cytomegaloviruses: Methods and Protocols

1. Overview of Human Cytomegalovirus Pathogenesis

            Heather L. Fulkerson, Maciej T. Nogalski, Donna Collins-McMillen, and Andrew D. Yurochko

 

2. Distinct Properties of Human Cytomegalovirus Strains and the Appropriate Choice of Strains for Particular Studies

            Giada Frascaroli and Christian Sinzger

 

3. Use of Diploid Human Fibroblasts as a Model System to Culture, Grow, and Study Human Cytomegalovirus Infection

            Elizabeth A. Fortunato

 

4. The Use of Primary Human Cells to Analyze Human Cytomegalovirus Biology

            Emma Poole, Ian Groves, Sarah Jackson, Mark Wills, and John Sinclair

 

5. Human Hematopoietic Long-Term Culture (hLTC) for Human Cytomegalovirus Latency and Reactivation

            Megan Peppenelli, Jason Buehler, and Felicia Goodrum

 

6. Collection and Isolation of CD14⁺ Primary Human Monocytes via Dual Density Centrifugation as a Model System to Study Human Cytomegalovirus Infection and Pathogenesis

            Bailey S. Mosher, Heather Fulkerson, and Andrew D. Yurochko

 

7. Stable and Inducible Gene Knockdown in Primary Human Fibroblasts: A Versatile Tool to Study the Role of Human Cytomegalovirus Host Cell Factors

            Anne-Charlotte Stilp, Patrick König, Myriam Scherer, and Thomas Stamminger

 

8. Construction of Human Cytomegalovirus Mutants with Markerless BAC Mutagenesis

            M. Zeeshan Chaudhry, Martin Messerle, and Luka Cicin-Sain

 

9. Methods for Studying the Function of Cytomegalovirus GPCRs

            Christine M. O'Connor and William E. Miller

 

10. Analysis of Cytomegalovirus Glycoprotein and Cellular Receptor Interactions

            Jamil Mahmud and Gary C. Chan

 

11. Antibody-Independent Quantification of Cytomegalovirus Virion Protein Incorporation Using HiBiT

            Iris K.A. Jones and Daniel N. Streblow

 

12. Use of a Phosphoproteomic Screen to Profile Early Changes during HCMV Infection of Human Monocytes

            Liudmila S. Chesnokova and Andrew D. Yurochko

 

13. A Generally Applicable CRISPR/Cas9 Screening Technique to Identify Host Genes Required for Virus Infection as Applied to Human Cytomegalovirus (HCMV) Infection of Epithelial Cells

            Xiaofei E and Timothy F. Kowalik

 

14. Quantitative Electron Microscopy to Study HCMV Morphogenesis

            Clarissa Read, Paul Walther, and Jens von Einem

 

15. Detection of Cytomegalovirus Interleukin 10 (cmvIL-10) by Enzyme-Linked Immunosorbent Assay (ELISA)

            Vivian P. Young, Margarette C. Mariano, Lionel Faure, and Juliet V. Spencer

 

16. Techniques for Characterizing Cytomegalovirus-Encoded miRNAs

            Nicole L. Diggins, Lindsey B. Crawford, Hillary M. Struthers, Lauren M. Hook, Igor Landais, Rebecca L. Skalsky, and Meaghan H. Hancock

 

17. Development of a huBLT Mouse Model to Study HCMV Latency, Reactivation, and Immune Response

            Lindsey B. Crawford and Patrizia Caposio

 

18. Rodent Models of Congenital Cytomegalovirus Infection

            Berislav Lisnic, Jelena Tomac, Djurdjica Cekinovic, Stipan Jonjic, and Vanda Juranic Lisnic

 

19. Recent Approaches and Strategies in the Generation of Anti-Human Cytomegalovirus Vaccines

            Suresh B. Boppana and William J. Britt