Hepatitis C Protocols

ISBN-13: 9781489943484 / Angielski / Miękka / 2013 / 621 str.

Johnson Y. N. Lau

Opis

The identification of hepatitis C virus by Michael Houghton and his c- leagues at the Chiron Corporation nearly years ago represented a technical tour de force of modern molecular medicine. This breakthrough not only unearthed the causative agent of non-A non-B hepatitis that had eluded the best of scientists for more than 20 years, but also was symbolic of another chapter in the changing paradigm of modern science and medicine. The re- lutionary concept of identifying a pathogen without actually visualizing or detecting it will forever redefine the way we approach pathogenesis of d- eases whose cause is unknown. It is benefitting that this discovery treads on the heels of the human immunodeficiency pandemic, and parallels the ala- ing emergence of various microbial pestilences in the world. Despite the rapid advances in our understanding of the virus, there is much remaining to learn about it. This book highlights some of the important areas yet to be unraveled. As illustrated in this book, the marriage between basic science and clinical medicine is essential in our quest for these unknowns. The last decade has focused on the fundamentals of the virus; the next decade must underscore translational research, bridging clinical medicine and basic science with re- ization of fundamental knowledge to improve prevention, diagnosis, and tre- ment of viral hepatitis C.

Recenzje

"Many leading researchers in the field are included in the book. . .It describes state-of-the-art methods and tools that are being used to address important medical and scientific questions. . .not only a highly practical manual for people working in laboratory medicine. . .also valuable for virologists, gastroenterologists, hepatologists and other medical infectious disease specialists."-Infection

"The useful clinical methods sections include description of methods for the detection of antibodies to HCV, quantitative HCV serum RNA assays, polymerase chain reaction (PCR) assays for the detection of HCV in liver and other tissues, HCV genotyping methods, and detection of HCV by in situ hybridization....Additional sections describe methods used by research, development, and production laboratories to clone and express HCV genes, to study viral replication and viral protein function, to produce recombinant HCV proteins, and to create in vitro culture models to allow study of viral replication and possible anitiviral compounds....The "Notes" sections will be particularly useful to individuals setting up the method for the first time. All the chapters contain current information and are well written....the scope of the book of use to anyone desiring to expand his or her knowledge of HCV. The introductory material for the sections is the best compilation of HCV-related information available in print today and makes Hepatitis C Protocols a valuable addition to the published literature on HCV."-Laboratory Medicine

"In summary Johnson Yiu-Nam Lau should be applauded for assembling an outstanding international ensemble of experts to produce a truly authoritative handbook for HCV clinicians and researchers."-The Quarterly Review of Biology

". . . an excellent review of the current and most up-to-date methods in molecular medicine involving hepatitis C. . . clinicians should review this textbook to give them a better understanding of the molecular techniques and shortcomings of the various diagnostic assays that they use in their day to day practice."-Canadian Journal of Infectious Disease

Spis treści

I. Detection of Anti-HCV in Serum Detection of Anti-GOR Antibodies in HCV Infection Confirmation of HCV Antibodies by the Line Immunoassay INNO-LIA HCV Ab III II. Detection and Quantitation of HCV in Serum Detection of HCV RNA in Serum by Reverse Transcriptase-PCR and Radiolabeled Liquid Hybridization Detection of HCV RNA in Serum by Reverse Transcription Polymerase Chain Reaction (RT-PCR) The Amplicor® HCV Tests for the Detection and Quantitation of Serum or Plasma HCV RNA Quantification of HCV RNA in Clinical Specimens by Branched DNA (bDNA) Technology Quality Control of the Polymerase Chain Reaction Preparation of Genotype-Specific HCV RNA Transcripts for Assessing HCV Detection and Quantification Assays III. Detection of HCV in Liver Tissue. Detection of HCV RNA in Formalin-Fixed, Paraffin-Embedded Liver Tissue by RT-PCR Quantification of HCV RNA in Liver Tissue by bDNA Assay IV. HCV Genotypes. Hepatitis C Virus: Types, Subtypes, and Beyond Molecular Evolutionary Analysis: Its Application in the Study of Hepatitis C Virus Genotyping by Type-Specific Primers That Can Type HCV Types 1-6 Genotyping Hepatitis C Virus by Type-Specific Primers for PCR Based on NS5 Region Determination of HCV Genotypes by RFLP HCV Genotyping by the Line Probe Assay INNO-LiPA HCV II Serological Genotyping Using Synthetic Peptides Derived from the NS4 Region Determination of HCV Quasispecies by Cloning and Sequencing Detection of Hepatitis C Virus Quasispecies Heterogeneity by Single-Strand Conformational Polymorphism Hepatitis C Virus Heteroduplex Tracking Assay: Application to Genotype Determination, Quasispecies Analysis, and Molecular Evolution Studies V. Detection of HCV In Situ. In Situ Detection of HCV: AnOverview In Situ Detection of Hepatitis C Viral Antigens In Situ Hybridization and the Detection and Localization of HCV RNA The In Situ Detection of PCR-Amplified Hepatitis C RNA. Immunoelectron Microscopic Characterization of HCV VI. Molecular Biologic Characterization. Cloning and Assembly of Complex Libraries of Full-Length HCV cDNA Clones Use of the Vaccinia Virus/T7 Expression System for Studying HCV Protein Processing Use of a Discistronic Vector for the Quantitation of HCV IRES Activity Expression and Dimerization of Hepatitis C Virus Core Protein in E. coli. Expression and Characterization of the HCV NS2 Protease Expression and Characterization of HCV NS3 Protease Expression and Characterization of the HCV NS3 Helicase Domain Hepatitis C Virus RNA-Dependent RNA Polymerase (NS5B Polymerase) Detection and Molecular Cloning of the Extreme 3'-End of HCV In Vitro Stability of Hepatitis C Virus RNA Methods for the Study of Sequence-Specific Binding of Proteins to the HCV RNA Genome VII. HCV-Specific Immunologic Response. Role of Immune Response in HCV Determination of Hepatitis C Virus-Specific CD4+ T-Cell Activity in PBMC Measurement of HCV-Specific CD8+ Cytotoxic T-Cell Activities in the Peripheral Blood by Europium Release Assay Determination of HCV-Specific Bulk CD8+ Activity in Liver Characterization of HCV-Specific Cytotoxic T-Lymphocytes from Liver Tissue Production of Human Monoclonal Antibodies to Hepatitis C Virus and Their Characterization VIII. In Vitro Culture Model. Specific Detection of Negative Strand RNA of Hepatitis C Virus Using Chemical RNA Modification Strand-Specific rTth RT-PCR for the Analysis of HCV Replication Cell Culture Systems for the Detection of HCV Infection

Blurb

In Hepatitis C Protocols, Johnson YN Lau and his expert collaborators present cutting-edge techniques for study of the hepatitis C virus infection and its disease, hepatitis C, which afflict an estimated 170 million people worldwide. The methods range from the detection of anti-HCV in serum and HCV in liver tissue, to HCV genotyping and the detection and quantification of HCV in serum. In addition to these key protocols, the authoritative contributors also evaluate recombinant proteins in immunological test systems, including the identification of important antigenic epitopes, antibodies, and recombinant antigens. Molecular and immunological techniques to study this virus and its infection, as well as
animal models and recombinant virus technology commonly used to express hepatitis C viral proteins, are also developed.

Hepatitis C Protocols summarizes the state-of-the-art in today's research and understanding of the virology of hepatitis C virus. The methods and systems detailed in this book will play a key role in the development of more effective antivirals, in the development of vaccines, and in a deeper understanding of viral replication strategies, viron structure, and the mechanisms of viron assembly and release. A valuable new standard resource for all researchers and clinicians, Hepatitis C Protocols leads the way
toward improved prevention, diagnosis, and treatment of viral hepatitis C.