MERS Coronavirus Methods and Protocols

This volume provides various techniques and methodologies currently used in the study of MERS-CoV. Chapters are divided into four parts detailing evolution and entry of MERS-coronavirus, genetic alteration and structural determination of MERS-coronavirus proteins, quantitation of virus and anti-viral factors, and mouse models for MERS -coronavirus.   Written in the highly successful Methods in Molecular Biology series 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 cutting-edge, MERS Coronavirus: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Studying Evolutionary Adaptation of MERS-CoV.- Evaluating MERS-CoV Entry Pathways.- Biochemical Characterization of Middle East Respiratory Syndrome Coronavirus Spike Protein Proteolytic Processing.- Crystallization and Structural Determination of the Receptor-binding Domain of MERS-CoV Spike Glycoprotein.- Bacterial Artificial Chromosome-based Lambda Red Recombination with the I-SceI Homing Endonuclease for Genetic Alteration of MERS-CoV.- Deducing the Crystal Structure of MERS-CoV Helicase.- Antigen Capture Enzyme-linked Immunosorbent Assay for Detecting Middle East Respiratory Syndrome Coronavirus in Humans.- Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR.- Evaluation of MERS-CoV Neutralizing Antibodies in Sera Using Live Virus Microneutralization Assay.- Generation of MERS-CoV Pseudotyped Viral Particles for the Evaluation of Neutralizing Antibodies in Mammalian Sera.- Qualitative and Quantitative Determination of MERS-CoV S1-specific Antibodies using ELISA.- Genetically Engineering a Susceptible Mouse Model for MERS-CoV-Induced Acute Respiratory Distress Syndrome.- Development of  a Mouse Adapted MERS Coronavirus.- Metabolite, Protein, and Lipid Extraction (MPLEx): A Method that Simultaneously Inactivates Middle East Respiratory Syndrome Coronavirus and Allows Analysis of Multiple Host Cell Components Following Infection.- Evaluation of Activation and Inflammatory Activity of Myeloid Cells DuringPathogenic Human Coronavirus Infection.- Histopathologic Evaluation and Scoring of Viral Lung Infection.