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Characterization of Poxvirus-Encoded Proteins that Regulate Innate Immune Signaling Pathways

Innate immune recognition of pathogens is critical to the prompt control of infections, permitting the host to survive to develop long-term immunity via an adaptive immune response. Poxviruses encode a family of proteins that inhibit signaling by Toll-like receptors to their downstre ...

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Application of Quartz Crystal Microbalance with Dissipation Monitoring Technology for Studying Interactions of Poxviral Proteins with Their Ligands

Poxviruses are one of the most complex of animal viruses and encode for over 150 proteins. The interactions of many of the poxviral-encoded proteins with host proteins, as well as with other proteins, such as transcription complexes, have been well characterized at the qualitative level. Some h ...

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Central Nervous System Distribution of the Poxviral Proteins After Intranasal Administration of Proteins and Titering of Vaccinia Virus in the Brain A

Poxviral proteins are known to interact with the immune system of the host. Some of them interact with the transcription factors of the host, whereas others interact with the components of the immune system. Vaccinia virus secretes a 28.8-kDa complement control protein (VCP), which is known to re ...

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Working Safely with Vaccinia Virus: Laboratory Technique and Review of Published Cases of Accidental Laboratory Infections

Vaccinia virus (VACV), the prototype orthopoxvirus, is widely used in the laboratory as a model system to study various aspects of viral biology and virus–host interactions, as a protein expression system, as a vaccine vector, and as an oncolytic agent. The ubiquitous use of VACVs in the laboratory ...

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In-Fusion Cloning with Vaccinia Virus DNA Polymerase

Vaccinia virus DNA polymerase (VVpol) encodes a 3′-to-5′ proofreading exonuclease that can degrade the ends of duplex DNA and expose single-stranded DNA tails. The reaction plays a critical role in promoting virus recombination in vivo because single-strand annealing reactions can th ...

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Genetic Manipulation of Poxviruses Using Bacterial Artificial Chromosome Recombineering

Traditional methods for genetic manipulation of poxviruses rely on low-frequency natural recombination in virus-infected cells. Although these powerful systems represent the technical foundation of current knowledge and applications of poxviruses, they require long (≥ ...

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Easy and Efficient Protocols for Working with Recombinant Vaccinia Virus MVA

Modified vaccinia virus Ankara (MVA) is a highly attenuated and replication-deficient strain of vaccinia virus that is increasingly used as vector for expression of recombinant genes in the research laboratory and in biomedicine for vaccine development. Major benefits of MVA inclu ...

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Screening for Vaccinia Virus Egress Inhibitors: Separation of IMV, IEV, and EEV

Concerns about the possible use of variola virus as a biological weapon as well as the need for therapeutics for the treatment or prevention of naturally acquired poxvirus infections or vaccination complications have led to the search for small molecule inhibitors of poxvirus replicati ...

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Isolation of Recombinant MVA Using F13L Selection

Modified vaccinia virus Ankara (MVA) has become a widely used vector for vaccine and laboratory purposes. Despite significant advances in recombinant MVA technology, the isolation of recombinant viruses remains a tedious and difficult process. This chapter describes the use of an eff ...

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Imaging of Vaccinia Virus Entry into HeLa Cells

The recently developed technique of live cell imaging has found numerous applications, including the detection of virus movements in living cells. To monitor virus motility, viruses or cellular proteins are fused with fluorescence markers and then detected by time-lapse fluoresce ...

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New Method for the Assessment of Molluscum Contagiosum Virus Infectivity

Molluscum contagiosum virus (MCV), a poxvirus pathogenic for humans, replicates well in human skin in vivo, but not in vitro in standard monolayer cell cultures. In order to determine the nature of the replication deficiency in vitro, the MCV infection process in standard culture has to be studied ...

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An Intradermal Model for Vaccinia Virus Pathogenesis in Mice

Intradermal injection of vaccinia virus in the ear pinnae of mice provides a model of dermal infection and vaccination. The key features of this model are the appearance of a lesion on the surface of the ear that can be measured as a clinical sign of disease and substantial growth of virus in the infected skin ...

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Protocols for Detection and Typing of Treponema pallidum Using PCR Methods

Syphilis, caused by Treponema pallidum, is a sexually transmitted disease which was epidemic in Europe between the 15th and 20th century. From 2000 onwards it is worldwide mostly encountered among men who have sex with men but also among women in resource poor setting. Syphilis can easily be treat ...

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Protocol for the Detection of Mycoplasma genitalium by PCR from Clinical Specimens and Subsequent Detection of Macrolide Resistance-Mediating Mutation

Mycoplasma genitalium is an established cause of male nongonococcal urethritis, in particular in cases with recurrent disease and in those negative for Chlamydia trachomatis. In women M. genitalium causes cervicitis and there is increasing evidence that it is causing pelvic inflam ...

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Validation of a Sensitive and Specific Real-Time PCR for Detection and Quantitation of Hepatitis B Virus Covalently Closed Circular DNA in Plasma of C

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) serves as a template for viral replication and plays a role in persistence of HBV infection. The origin and significance of cccDNA in plasma, however, are not well understood. A sensitive, specific, and reproducible real-time PCR ...

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The Molecular Diagnosis of Sexually Transmitted Genital Ulcer Disease

Highly sensitive and specific nucleic acid amplification tests (NAATs) have emerged as the gold standard diagnostic tests for many infectious diseases. Real-time PCR has further refined the technology of nucleic acid amplification with detection in a closed system and enabled mult ...

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Guidelines for the Use of Molecular Tests for the Detection and Genotyping of Human Papilloma Virus from Clinical Specimens

Accurate genotyping of a human papilloma virus (HPV) isolated from clinical specimens depends on molecular identification of the unique and exclusive nucleotide base sequence in the hypervariable region of a highly conserved segment of the HPV L1 gene. Among other options, a heminested ...

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Guidelines for High-Resolution Genotyping of Chlamydia trachomatis Using Multilocus Sequence Analysis

Chlamydia is one of the most common sexually transmitted infections worldwide and can cause ectopic pregnancies and infertility. It is therefore important to have adequate genotyping tools for investigating the spread of Chlamydia trachomatis among the population. Here, we descr ...

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Guidelines for the Quantification of HIV and HCV in Small Volume Whole Blood Samples

The quantification of Hepatitis C Virus (HCV) and Human Immunodeficiency Virus (HIV) in whole blood provides several advantages over the quantification in plasma samples. The application of small samples of capillary blood allows for application in point-of-care diagnostic test ...

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Guidelines for the Qualitative Detection of Viral Genomes in Dried Blood Spots

Dried blood spots (DBSs) are a useful alternative to blood sampling especially in children or for screening high-risk populations in developing countries. DBS blood collection can be employed in the diagnosis of viral infections by PCR or RT-PCR and also in viral genome sequencing. In additi ...

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