Springer protocols feed by Infectious Diseases 
This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.
This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Infectious Viral Quantification of Chikungunya Virus & mdash;Virus Plaque Assay
The plaque assay is an essential method for quantification of infectious virus titer. Cells infected with virus particles are overlaid with a viscous substrate. A suitable incubation period results in the formation of plaques, which can be fixed and stained for visualization. Here, we describe a method for measuring Chikungunya virus (CHIKV) titers via virus plaque assays. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Bioinformatics Based Approaches to Study Virus & ndash;Host Interactions During Chikungunya Virus Infection
The limitations of high-throughput genomic methods used for studying virus & ndash;host interactions make it difficult to directly obtain insights on virus pathogenesis. In this chapter, the central steps of a protein structure similarity based computational approach used to predict the host interactors of Chikungunya virus are explained by highlighting the important aspects that need to be considered. Identification of such conserved set of putative interactions that allow the virus to take control of the host has the potential to deepen our understanding of the virus-specific remodeling processes of the host cell and ill...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Viral & ndash;Host Protein Interaction Studies Using Yeast Two-Hybrid Screening Method
Yeast two-hybrid (Y2H) assay is one of the earliest methods developed to study protein & ndash;protein interactions. In the proteomics era, Y2H has created a niche of its own by providing protein interaction maps for various organisms. Owing to limited coding capacities of their genomes, viruses are dependent on their host cellular machinery for successful infection. Identification of the key players orchestrating the survival of virus in their host is essential for understanding viral life cycle and devising strategies to prevent interactions resulting in pathogenesis. In this chapter, Y2H assay will be explained in detai...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Neutralization Assay for Chikungunya Virus Infection: Plaque Reduction Neutralization Test
Neutralization assay is a technique that detects and quantifies neutralizing antibody in serum samples by calculating the percentage of reduction of virus activity, as the concentration of virus used is usually constant. Neutralizing antibody titer is conventionally determined by calculating the percentage reduction in total virus infectivity by counting and comparing number of plaques (localized area of infection due to cytopathic effect) with a standard amount of virus. Conventional neutralizing test uses plaque-reduction neutralization test (PRNT) to determine neutralizing antibody titers against Chikungunya virus (CHIK...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Using Bicistronic Baculovirus Expression Vector System to Screen the Compounds That Interfere with the Infection of Chikungunya Virus
Chikungunya virus (CHIKV) is the etiologic agent of Chikungunya fever and has emerged in many countries over the past decade. There are no effective drugs for controlling the disease. A bicistronic baculovirus expression system was utilized to co-express CHIKV structural proteins C (capsid), E2 and E1 and the enhanced green fluorescence protein (EGFP) in Spodoptera frugiperda insect cells (Sf21). The EGFP-positive Sf21 cells fused with each other and with uninfected cells to form a syncytium is mediated by the CHIKV E1 allowing it to identify chemicals that can prevent syncytium formation. The compounds characterized by th...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
A Real-Time Cell Analyzing Assay for Identification of Novel Antiviral Compounds against Chikungunya Virus
Screening of viral inhibitors through induction of cytopathic effects (CPE) by conventional method has been applied for various viruses including Chikungunya virus (CHIKV), a significant arbovirus. However, it does not provide the information about cytopathic effect from the beginning and throughout the course of virus replication. Conventionally, most of the approaches are constructed on laborious end-point assays which are not capable for detecting minute and rapid changes in cellular morphology. Therefore, we developed a label-free and dynamical method for monitoring the cellular features that comprises cell attachment,...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Antiviral Strategies Against Chikungunya Virus
In the last few decades the Chikungunya virus (CHIKV) has evolved from a geographically isolated pathogen to a virus that is widespread in many parts of Africa, Asia and recently also in Central- and South-America. Although CHIKV infections are rarely fatal, the disease can evolve into a chronic stage, which is characterized by persisting polyarthralgia and joint stiffness. This chronic CHIKV infection can severely incapacitate patients for weeks up to several years after the initial infection. Despite the burden of CHIKV infections, no vaccine or antivirals are available yet. The current therapy is therefore only symptoma...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Immunohistochemical Detection of Chikungunya Virus Antigens in Formalin-Fixed and Paraffin-Embedded Tissues
Immunohistochemistry is a histological technique that allows detection of one or more proteins of interest within a cell using specific antibody binding, followed by microscopic visualization of a chromogenic substrate catalyzed by peroxidase and/or alkaline phosphatase. Here, we describe a method to localize Chikungunya virus (CHIKV) antigens in formalin-fixed and paraffin-embedded infected mouse brain. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Generation of Mouse Monoclonal Antibodies Specific to Chikungunya Virus Using ClonaCell-HY Hybridoma Cloning Kit
Monoclonal antibodies offer high specificity and this makes it an important tool for molecular biology, biochemistry and medicine. Typically, monoclonal antibodies are generated by fusing mouse spleen cells that have been immunized with the desired antigen with myeloma cells to create immortalized hybridomas. Here, we describe the generation of monoclonal antibodies that are specific to Chikungunya virus using ClonaCell-HY system. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Mouse Models of Chikungunya Virus
The majority of medical advances have been made using animals. Studies using mouse models of chikungunya-induced disease have proven invaluable for dissecting the intricate nature of the immune response to this viral infection and identifying potential targets for the development of treatment strategies. Herein we describe the common mouse models used to research the pathobiology of chikungunya virus infection to date. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Infectious Viral Quantification of Chikungunya Virus—Virus Plaque Assay
The plaque assay is an essential method for quantification of infectious virus titer. Cells infected with virus particles are overlaid with a viscous substrate. A suitable incubation period results in the formation of plaques, which can be fixed and stained for visualization. Here, we describe a method for measuring Chikungunya virus (CHIKV) titers via virus plaque assays. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Propagation of Chikungunya Virus Using Mosquito Cells
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that transmits in between a mosquito host vector to a primate host and then back to the mosquito host vector to complete its life cycle. Hence, CHIKV must be able to replicate in both host cellular systems that are genetically and biochemically distinct. The ability to grow and propagate the virus in high titers in the laboratory is fundamentally crucial in order to understand virus replication in different host cellular systems and many other CHIKV research areas. Here, we describe a method on CHIKV propagation using C6/36, a mosquito cell line derived from Aedes al...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
T-Cell Epitope Prediction of Chikungunya Virus
There has been a growing demand for vaccines against Chikungunya virus (CHIKV), and epitope-based vaccine is a promising solution. Identification of CHIKV T-cell epitopes is critical to ensure successful trigger of immune response for epitope-based vaccine design. Bioinformatics tools are able to significantly reduce time and effort in this process by systematically scanning for immunogenic peptides in CHIKV proteins. This chapter provides the steps in utilizing machine learning algorithms to train on major histocompatibility complex (MHC) class I peptide binding data and build prediction models for the classification of b...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Bioinformatics Based Approaches to Study Virus–Host Interactions During Chikungunya Virus Infection
The limitations of high-throughput genomic methods used for studying virus–host interactions make it difficult to directly obtain insights on virus pathogenesis. In this chapter, the central steps of a protein structure similarity based computational approach used to predict the host interactors of Chikungunya virus are explained by highlighting the important aspects that need to be considered. Identification of such conserved set of putative interactions that allow the virus to take control of the host has the potential to deepen our understanding of the virus-specific remodeling processes of the host cell and illum...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
Application of GelC-MS/MS to Proteomic Profiling of Chikungunya Virus Infection: Preparation of Peptides for Analysis
Gel-enhanced liquid chromatography coupled with tandem mass spectrometry (GeLC-MS/MS) is a labor intensive, but relatively straightforward methodology that generates high proteome coverage which can be applied to the proteome analysis of a range of starting materials such as cells or patient specimens. Sample proteins are resolved electrophoretically in one dimension through a sodium dodecyl sulfate (SDS) polyacrylamide gel after which the lanes are sliced into sections. The sections are further diced and the gel cubes generated are subjected to in-gel tryptic digestion. The resultant peptides can then be analyzed by tande...
Source: Springer protocols feed by Infectious Diseases - December 31, 2015 Category: Infectious Diseases Source Type: news
