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.

Inhalt
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