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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.
List of contents
Overview of Human Cytomegalovirus Pathogenesis.- Distinct Properties of Human Cytomegalovirus Strains and the Appropriate Choice of Strains for Particular Studies.- Use of Diploid Human Fibroblasts as a Model System to Culture, Grow, and Study Human Cytomegalovirus Infection.- The Use of Primary Human Cells to Analyze Human Cytomegalovirus Biology.- Human Hematopoietic Long-Term Culture (hLTC) for Human Cytomegalovirus Latency and Reactivation.- Collection and Isolation of CD14+ Primary Human Monocytes via Dual Density Centrifugation as a Model System to Study Human Cytomegalovirus Infection and Pathogenesis.- Stable and Inducible Gene Knockdown in Primary Human Fibroblasts: A Versatile Tool to Study the Role of Human Cytomegalovirus Host Cell Factors.- Construction of Human Cytomegalovirus Mutants with Markerless BAC Mutagenesis.- Methods for Studying the Function of Cytomegalovirus GPCRs.- Analysis of Cytomegalovirus Glycoprotein and Cellular Receptor Interactions.- Antibody-Independent Quantification of Cytomegalovirus Virion Protein Incorporation Using HiBiT.- Use of a Phosphoproteomic Screen to Profile Early Changes during HCMV Infection of Human Monocytes.- 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.- Quantitative Electron Microscopy to Study HCMV Morphogenesis.- Detection of Cytomegalovirus Interleukin 10 (cmvIL-10) by Enzyme-Linked Immunosorbent Assay (ELISA).- Techniques for Characterizing Cytomegalovirus-Encoded miRNAs.- Development of a huBLT Mouse Model to Study HCMV Latency, Reactivation, and Immune Response.- Rodent Models of Congenital Cytomegalovirus Infection.- Recent Approaches and Strategies in the Generation of Anti-Human Cytomegalovirus Vaccines.
