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
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.- 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.- 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.- HCV Genotypes.- Hepatitis C Virus.- Molecular Evolutionary Analysis.- Genotyping by Type-Specific Primers That Can Type HCV Types 16.- 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.- Detection of HCV In Situ.- In Situ Detection of HCV.- 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.- lmmunoelectron Microscopic Characterization ofHCV.- 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.- 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.- 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.- Replication and Detection of Hepatitis C Virus in Liver-Derived Cell Lines.- Primary Human Hepatocyte Culture for the Study of HCV.- A Cultivation Method for Highly Differentiated Primary Chimpanzee Hepatocytes Permissive for Hepatitis C Virus Replication.- Construction ofRecombinant Viral Vectors for the Expression of HCV Genes.- Construction of Recombinant Vaccinia Virus Expressing HCV Genes.- Construction of Recombinant Adeno-Associated Virus (AAV).- Production of Replication-Deficient Adenovirus Recombinants.- Generation of Recombinant Herpes Simplex Virus Ampiicons.- Construction of Recombinant Sindbis-Based Expression Vectors for the Study of HCV Genes and Their Products.- In Vivo Models.- The Chimpanzee Model.- Liver Transplantation as a Model to Study Hepatitis C Virus Infection.