The polymerase chain reaction (PCR) has revolutionized both the basic and the applied aspects of the biomedical field with more than 40,000 papers having been published employing this technique. PCR has become an indispensable tool for basic research applications, such as cloning (1), seq ...

Hepatitis C virus (HCV) RNA has been detected in the sera and liver sections of patients with chronic HCV infection using RT-PCR and other sensitive molecular techniques. Since archived formalin-fixed, paraffin embedded tissues from these patients are readily available for retrospe ...

With this statement, Sherlock and Dooley have described two of the three major challenges involved in quantitatively measuring any analyte in tissue samples: the distribution of the analyte in the tissue; and the standard of reference, or denominator, with which to make comparisons betwe ...

Hepatitis C virus (HCV), the etiological agent responsible for the majority of cases of parenterally acquired liver disease, is found throughout the world. HCV is an enveloped virus with a small, single-stranded RNA genome. Because it uses an error-prone, RNA-dependent RNA polymerase, HCV h ...

Non-A, non-B hepatitis was recognized as a frequent consequence of blood transfusion for many years before the agent responsible, hepatitis C virus (HCV), was first cloned and sequenced in 1989 Very quickly it became apparent that viruses from different parts of the world were distinct, and aft ...

The recent development of high-power personal computer hardware and software has allowed investigators to prepare phylogenetic trees without understanding of the fundamental principles of molecular evolution. Unfortunately, inexperienced investigators are also pr ...

Hepatitis C virus (HCV) was identified as a major causative agent of non-A, non-B hepatitis (1). Numerous complete or partial nucleotide sequences of HCV isolates have been reported worldwide, and comparison of these sequences revealed their marked genetic heterogeneity nature, sugge ...

Hepatitis C virus (HCV), a flavi-like virus with a positive-sense, single-stranded genome RNA (1–3), shows considerable variation in nucleotide sequences. These variations fall into a series of specific patterns and are the basis for classification of HCV into different types and genoty ...

Several different methods have been developed for the typing of HCV variants: direct sequence analysis, slot-blot hybridization analysis of reverse transcriptase-polymerase chain reaction (RT-PCR) products using cDNA probes specific to each HCV genotype and PCR amplificati ...

Hepatitis C viruses (HCVs) constitute a highly variable genus within the Flaviviridae, with closest homology to the hepatitis G and GB viruses, and Pestiviruses. The positive-stranded RNA genome encodes a polyprotein which is co- and posttranslationally cleaved into at least nine prot ...

The RNA genome of hepatitis C virus (HCV) displays extensive sequence variation, and consequently, the virus is classified into six major genotypes. The severity of disease and response to antiviral treatment are thought to be influenced by both viral and host-related factors, including a ...

Most RNA viruses exists as a heterogeneous mixture of closely related viral genome in the host, which is the result of high error rates in RNA replication and selected by various viral and host factors. The spectrum of this spectrum of related genomes within a host is referred as quasispecies (1). This im ...

The heteroduplex tracking assay (HTA) is a tool that can be used for determining genotype, quasispecies analysis, molecular evolution, and epidemiological studies (1–7). By hybridizing a labeled, single-stranded DNA probe to colinear, reverse transcriptase (RT) PCR products from a s ...

Hepatitis C virus (HCV) is believed to replicate via a viral-encoded, RNA-dependent RNA polymerase. This replication strategy has limited fidelity Thus, HCV is genetically heterogenous. To date, six HCV genotypes and more than 80 viral subtypes have been identified. Further, even within i ...

Hepatitis C virus (HCV) is the agent responsible for the majority of cases of the parenterally transmitted non-A, non-B hepatitis. The major obstacles to its discovery were the low level of replication in the infected host, both natural and experimental, and the low immunogenicity of its protei ...

A reliable detection system for HCV antigens in liver tissue may be used to identify the HCV cellular tropism and subcellular sites of viral replication Also, it can be used to study the relationship between viral expression and disease activity. Finally, it can facilitate the study of host-viral ...

The procedure described below was originally reported to detect the hepatitis C virus RNA (genomic strand) by nonradioisotopic in situ hybridization in formalin-fixed, paraffin-embedded liver tissue of two acutely infected chimpanzees, in a collaborative study conducted with R. ...

The isolation of plasmid DNA from bacteria is a crucial technique in molecular biology and is an essential step in many procedures such as cloning, DNA sequencing, transfection, and gene therapy. These manipulations require the isolation of high-purity plasmid DNA. Commercial anion-ex ...

Plasmid extraction is typically performed to produce template DNA for a desired molecular biological reaction, or set of reactions, such as restriction endonuclease digestion (see Chapter 20), DNA sequencing (see Chapter 22), in vitro mutagenesis (see Chapters 23–26), transformat ...

Cosmid and bacterial artificial chromosome (BAC) systems have been developed for the cloning of large DNA inserts averaging 40 kb and 130 kb (range: 90–300 kb), respectively. The resulting clones are more stable than yeast artificial chromosomes (YACs) and rarely chimeric, which makes them e ...