Hepatitis C

By providing a compilation of cutting-edge research techniques currently used in HCV labs worldwide, this book fully updates the previous edition. It reflects the progress which promises to unlock the remaining vital information regarding this human pathogen.

When I was ?rst approached by Dr. Hengli Tang about writing the foreword to this book, the second edition of Hepatitis C Virus, I was a little puzzled: Book forewords are penned by persons far more famous than I am. Surely there were better people to write this, with a handful of the true visionaries in the ?eld immediately coming to my mind. However, after some thought and self-re?ection, I realized that I might just be the right person for the job. I remember the publication of the ?rst edition of Hepatitis C Virus in 1998, the year I began reading about hepatitis C virus with great interest. I purchased a copy of the ?rst edition from my meager graduate student stipend and read it from cover to cover, along with a variety of hepatitis C publications, to try to get a feel if this hepatitis C virus ?eld was something I might want to work on in the future. At the time, the ?rst edition was among the most comprehensive collection of hepatitis C protocols and methods available in one volume. I was amazed at the progress that had been made in hepatitis C research, and even more amazed as I learned more about the experimental challenges facing the ?eld at that time.

Provides the most up-to-date technical information on the new cell culture infection system as written by leading experts in the field Presents both an overview and detailed protocols on the replicon system used by virtually all the HCV labs around the world Covers a wide range of multi-discipline approaches adopted for HCV research, including NMR-based structure determination and mathematical modelling Broadly applicable guides valuable for studying the replication of other positive-strand RNA viruses Includes supplementary material: sn.pub/extras

Klappentext

With the development of robust cell-culturing systems capable of Hepatitis C Virus replication and infection, the past decade has witnessed an explosion of innovative research directed at understanding HCV biology in the context of the authentic replicative cycle of the virus. By providing a compilation of cutting-edge research techniques currently in use in HCV labs worldwide, Hepatitis C: Methods and Protocols, Second Edition fully updates the previous edition to reflect this progress which promises to unlock the remaining vital information regarding this human pathogen and its interactions. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Fully updated and comprehensive, Hepatitis C: Methods and Protocols, Second Edition serves as a valuable reference guide for basic and clinical researchers using HCV as a model and striving to better understand this critical disease.

Inhalt
Detection and Quantification of HCV.- Detection and Quantification of Serum or Plasma HCV RNA: Mini Review of Commercially Available Assays.- Simultaneous Detection of Anti-HCV Antibody and HCV Core Antigen.- Immunohistochemical Detection of HCV Proteins in Liver Tissue.- HCV Nomenclature, Genotyping, and Quasispecies.- Nomenclature and Numbering of the Hepatitis C Virus.- A Real-Time Taqman® Method for Hepatitis C Virus Genotyping and Methods for Further Subtyping of Isolates.- Characterization of HCV Quasispecies in the Hypervariable Region 1 (HVR 1) by In Vitro Translation and Mass Spectrometry.- HCV Replication in Vitro.- Purification and Crystallization of NS5A Domain I of Hepatitis C Virus.- NS5A Phosphorylation and Hyperphosphorylation.- Preparation and Handling of Hepatitis C Viral Proteins NS3 and NS5B for Structural Studies.- Structural and Functional Analysis of the HCV p7 Protein.- HCV Replicons: Overview and Basic Protocols.- Reverse Transcription PCR-Based Sequence Analysis of Hepatitis C Virus Replicon RNA.- Studying HCV RNA Synthesis In Vitro with Replication Complexes.- Proteomics Study of the Hepatitis C Virus Replication Complex.- Investigation of the Hepatitis C Virus Replication Complex.- Regulation of Interferon Regulatory Factor 3-Dependent Innate Immunity by the HCV NS3/4A Protease.- Selection and Characterization of Drug-Resistant HCV Replicons In Vitro with a Flow Cytometry-Based Assay.- High-Throughput Screening of HCV RNA Replication Inhibitors by Means of a Reporter Replicon System.- Inhibition of HCV Replication by Small Interfering RNA.- HCV Entry and Infection.- Cellular Receptors and HCV Entry.- Studying HCV Cell Entry with HCV Pseudoparticles (HCVpp).- Screening of Small-Molecule Compounds as Inhibitors of HCV Entry.- Isolation ofJFH-1 Strain and Development of an HCV Infection System.- Measuring HCV Infectivity Produced in Cell Culture and In Vivo.- Genotype 1a HCV (H77S) Infection System.- Full-Length Infectious HCV Chimeras.- Adaptation of the Hepatitis C Virus to Cell Culture.- Primary Human Hepatocyte Culture for HCV Study.- In Vivo Study of HCV in Mice with Chimeric Human Livers.- Immunity and Modeling.- Determination of HCV-Specific T-Cell Activity.- Analysis of HCV-Specific T Cells by Flow Cytometry.- Detection of Neutralizing Antibodies with HCV Pseudoparticles (HCVpp).- Mathematical Modeling of HCV Infection and Treatment.