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Internal Transcribed Spacer (ITS)-PCR Identification of MRSA

Polymerase chain reaction (PCR) analysis of the 16S-23S rRNA gene internal transcribed spacer (ITS) followed by microchip gel electrophoresis was useful for identification of staphylococci and for strain delineation of Staphylococcus aureus. In the study presented in this chapt ...

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Multilocus Sequence Typing (MLST) of Staphylococcus aureus

Multilocus sequence typing (MLST) is a widely accepted method of DNA sequence-based typing that relies on analysis of relatively conserved genes that encode essential proteins. For Staphylococcus aureus, the level of discrimination provided by MLST is sufficient to provide a relati ...

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Staphylococcal Cassette Chromosome mec (SCCmec) Analysis of MRSA

Methicillin-susceptible Staphylococcus aureus changes to methicillin-resistant S. aureus (MRSA) on acquisition of the staphylococcal cassette chromosome mec (SCCmec). At least five types of SCCmec elements have been reported. All the SCCmec elements share four common chara ...

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Molecular Analysis of Staphylococcal Superantigens

Staphylococcal superantigens (SAgs) comprise a large family of exotoxins produced by Staphylococcus aureus strains. These exotoxins are important in a variety of serious human diseases, including menstrual and nonmenstrual toxic shock syndrome (TSS), staphylococcal pneu ...

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Targeted Gene Disruption for the Analysis of Virulence of Staphylococcus aureus 1

In recent years, molecular genetic approaches to the study of the disease pathogenesis of Staphylococcus aureus have resulted in many new biological insights. I describe methods used for targeted disruption of staphylococcal genes leading to loss of gene function, important for stud ...

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Cloning Single-Chain Antibody Fragments (scFv) from Hybridoma Cells

Despite the availability of antibody libraries for the selection of receptor molecules, the large number of established and well-characterized hybridoma lines still represent a useful source for recombinant antibody genes. This protocol describes the PCR amplification, clon ...

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Molecular Diagnostics Resources on the Internet

Busy diagnosticians need to know what is useful, and what is dross, when dealing with the internet. From the comprehensive array of resources that characterizes the offerings available via the world wide web and email correspondence, in particular, this chapter seeks to identify the most use ...

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Basic Problems of Serological Laboratory Diagnosis

Serological laboratory diagnosis of infectious diseases is inflicted with several kinds of basic problems. One difficulty relates to the fact that the serological diagnosis of infectious diseases is double indirect: The first indirect aim in diagnosing an infectious disease is to id ...

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Design and Preparation of Recombinant Antigens as Diagnostic Reagents in Solid-Phase Immunosorbent Assays

Analysis of the humoral immune response to infectious diseases has played, and will to continue to play, a key role in their diagnosis and immune surveillance. Although rapid genome detection methodologies, such as PCR, are beginning to replace immune assays for disease diagnosis, they are not ...

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Solid Supports in Enzyme-Linked Immunosorbent Assay and Other Solid-Phase Immunoassays

Most modern immunoassays involve the use of synthetic solid phases to immobilize one of the reactants, often by simple adsorption. These solid-phase immunoassays (SPIs) involve ligand-receptor interactions that occur within a reaction volume close to the solution/solid-phase i ...

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Propagation, Purification, and In Vivo Testing of Oncolytic Vesicular Stomatitis Virus Strains

Oncolytic viruses are self-amplifying therapeutics that specifically replicate in and kill cancer cells. We have previously shown that vesicular stomatitis virus (VSV) can be used as an oncolytic virus. A strain of VSV harboring a mutation in the M protein (VSVΔ51) was found to exhibit enhan ...

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Oncolytic Measles Virus Retargeting by Ligand Display

Despite significant advances in recent years, treatment of metastatic malignancies remains a significant challenge. There is an urgent need for development of novel therapeutic approaches. Virotherapy approaches have considerable potential, and among them measles virus (M ...

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Exploring Host Factors that Impact Reovirus Replication, Dissemination, and Reovirus-Induced Cell Death in Cancer Versus Normal Cells in Culture

Oncolytic viruses, such as reovirus, offer a promising approach to cancer treatment. Concurrently, oncolytic viruses provide a valuable tool for deciphering unique attributes of cancer cells that support superior virus replication, cell death, or virus dissemination. Through o ...

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Analysis of Three Properties of Newcastle Disease Virus for Fighting Cancer: Tumor-Selective Replication, Antitumor Cytotoxicity, and Immunostimulatio

Newcastle disease virus (NDV), a bird paramyxovirus, is an antitumor agent which has shown benefits to cancer patients. Its antineoplastic efficacy appears to be associated with three properties of the virus: 1.

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Next-Generation Oncolytic Vaccinia Vectors

Oncolytic vaccinia viruses have made some impressive advances over the last 5 years, with a range of �different backbones displaying significant antitumor responses in preclinical models, and some exciting clinical results being reported against liver cancers. Because the virus ...

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Evaluation of Innate Immune Signaling Pathways in Transformed Cells

Oncolytic viruses, the use of viruses to treat cancer, is emerging as a new option for cancer therapy. Oncolytic viruses, of both DNA and RNA origin, exhibit the ability to preferentially replicate in and kill cancer cells plausibly due to defects in innate immune signaling or translation regula ...

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Oncolytic Herpes Simplex Virus Engineering and Preparation

Herpes simplex virus-1 (HSV-1) is an enveloped, double-stranded DNA virus that has been used with modification as an oncolytic virus against a number of tumor types. Modifications that make HSV-1 replication-�conditional, i.e., selectively divide in replicating cells make it fulfill a pr ...

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Construction of Capsid-Modified Adenoviruses by Recombination in Yeast and Purification by Iodixanol-Gradient

Adenovirus represents a valuable tool for the treatment of cancer, but tumor targeting remains a pending issue. Most common procedures to modify adenovirus genome are time-consuming due to the requirement of multiple cloning steps, and the low efficacy of the recombination process. Here, ...

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Construction of Targeted and Armed Oncolytic Adenoviruses

Oncolytic (replication-competent) adenoviruses (Ads) represent the most advanced platform for cancer gene therapy. These viral vectors ablate tumors by killing tumor cells in the process of virus replication. As progeny virions are released, they infect remaining cancer cells, g ...

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Syrian Hamster Tumor Model to Study Oncolytic Ad5-Based Vectors

Oncolytic (replicating) adenovirus (Ad) vectors are emerging as a promising form of a cancer therapy agent. There has been a need for an appropriate animal model to study oncolytic Ad since human Ad �replication is usually species specific. We have shown that Syrian (golden) hamsters are an appr ...

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