Vaccinia, Mpox, and Other Poxviruses Methods and Protocols

This detailed volume covers new and classic methods for vaccinia virus (the prototype poxvirus) but also extends to methods and protocols specifically tailored for mpox virus, lumpy skin disease virus, and myxoma virus. As these viruses continue to expand their territories, research into the impact of poxviruses on human and animal health, as well as investigating vaccine vectors and oncolytic virotherapy, becomes more important, and this book provides a thorough resource for this area of study. 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, Vaccinia, Mpox, and Other Poxviruses: Methods and Protocols serves as an ideal resource for virologists working in this vital section of the field.

Poxvirus Epidemiology.- Mpox Specimen Collection, Storage, Preparation, and Laboratory Diagnostics by PCR.- A Quantitative Real-Time PCR Assay for Measuring Poxvirus Replication and Cell Binding.- Orthopoxvirus Genome Sequencing, Assembly, and Analysis.- Bioinformatics for the Structural Genomics of Poxviruses.- Construction and Isolation of Recombinant Vaccinia Virus by Homologous Recombination Using Fluorescent Protein Markers.- Rapid Generation of Recombinant Poxviruses Using CRISPR/Cas9 Gene Editing.- Cas9-Mediated Poxvirus Recombinant Recovery (CASPRR) for Fast Recovery of Recombinant Vaccinia Viruses.- Use of a Poxvirus K3 Ortholog as a Positive Selection Marker for Construction of Recombinant Vaccinia Virus with Modified Host Range.- Orthopoxvirus Purification for High Containment Laboratories.- Evaluation of Vaccinia Virus Infection in Mice Using Two-Reporter Recombinant Virus.- Bioluminescence Imaging to Study Recombinant Orthopoxvirus Infection in Animal Models.- Methods to Study Poxvirus Structures by Cryo-EM Imaging Modalities.- Investigating the Impact of Host Factors on Vaccinia Virus Infection through Single-Cell Analysis via Flow Cytometry.- High-Throughput Single Cell Analysis of Vaccinia Virus Infection.- Experimental Evolution of Poxviruses.- An Evolutionary Framework Exploiting Virologs and Their Host Origins to Inform Poxvirus Protein Functions.- Sample Preparation for Global Metabolic Profiling of Vaccinia Virus-Infected Primary Human Foreskin Fibroblasts.- Modified Vaccinia Virus Ankara Amplification and Titration Using Crystal Violet- or Immuno-Staining in DF-1 Cells.- Generating MVA-Vector Vaccine Candidates and Testing Them in Animal Models.- Isolation and Identification of Lumpy Skin Disease Virus.- Testing Oncolytic Myxoma Virus in Immunocompetent Mouse Model for Cancer Therapy.