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疾病及处理

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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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Sexually Transmitted Diseases: Reflections on Metaphors and Ethics

Sexually transmitted diseases (STDs) are probably the most tabooed diseases we know. The taboos and the related stigmata shape patients reality and influence significantly health care policies, medical research, and actual problems in medical ethics. To better understand these co ...

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Self-Collection of Specimens for Nucleic Acid-Based Diagnosis of Pharyngeal, Cervicovaginal, Urethral, and Rectal Neisseria gonorrhoeae and Chlamydia

Commercially available nucleic acid amplification tests (NAATs) for Neisseria gonorrhoeae and Chlamydia trachomatis detection allow for self-collection including home-based collection from multiple anatomic sites such as the urethra, cervicovagina, rectum, and phar ...

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Protocol for a Mammalian Cell-Based Assay for Monitoring the HIV-1 Protease Activity

Proteases are essential at different stages of the viral life cycle and for the establishment of a successful infection. Monitoring the catalytic activity of proteases in an easy and straightforward manner can thus drastically facilitate the discovery of novel antivirals, as well as help ...

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Protocol for the Clonal Analysis of NK Cell Effector Functions by Multi-parameter Flow Cytometry

Natural killer (NK) cells provide a first line of defense against viral infections and prepare the ground for subsequent action of virus-specific T cells in a concerted way. Human NK cells use a sophisticated system of inhibitory and stimulatory receptors of the killer cell immunoglobulin- ...

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Protocol for Analyzing Human Leukocyte Antigen Variants and Sexually Transmitted Infections: From Genotyping to Immunoassays

This chapter describes experimental and analytical procedures that can be used to decipher the specific role of human leukocyte antigen (HLA) variants in infectious diseases. The techniques are distilled from more than one decade of active immunogenetics research, primarily on sex ...

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Protocol for Gene Expression Profiling Using DNA Microarrays in Neisseria gonorrhoeae

Gene expression profiling using DNA microarrays has become commonplace in current molecular biology practices, and has dramatically enhanced our understanding of the biology of Neisseria spp., and the interaction of these organisms with the host. With the choice of microarray platf ...

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