Bibliographic Information

Hepatitis C protocols

edited by Johnson Yiu-Nam Lau

(Methods in molecular medicine / John M. Walker, series editor, 19)

Humana Press, c1998

Available at  / 5 libraries

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Note

Forewords by T. Jake Liang, Gary L. Davis, Michael Houghton, Q.L. Choo, and George Kuo

Includes bibliographical references and index

Description and Table of Contents

Description

Johnson Lau and his expert collaborators present cutting-edge techniques for study of the hepatitis C virus, a disease afflicting over 100 million people worldwide. The methods range from the detection of anti-HCV in serum and HCV in liver tissue to HCV genotypes 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. The methods and systems detailed in this book will play a key role in the development of more effective antivirals, in the introduction of vaccines, and in a deeper understanding of viral replication strategies, viron structure, and the mechanisms of viron assembly and release.

Table of Contents

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(R) 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

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