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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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Extraction of DNA from Dried Blood in the Diagnosis of Congenital CMV Infection

Viral DNA detection in dried blood spotted on filter paper, dried blood spots (DBS), is valuable in the diagnosis of viral infections, with at the moment congenital cytomegalovirus (CMV) being the most common application. CMV detection in clinical samples taken within the first 2–3 weeks after ...

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Protocol for the Use of a Bead Array for the Multiple Detection of Genotype of Chlamydia trachomatis

The identification of Chlamydia trachomatis genotypes is important for both molecular epidemiology and infection control such as contact tracing and identification of high-risk groups. Currently, at least 19 human serovars have been recognized by using polyclonal and monoclo ...

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Protocol for the Use of PCR-Denaturing Gradient Gel Electrophoresis and Quantitative PCR to Determine Vaginal Microflora Constitution and Pathogens in

In healthy women, the vaginal ecosystem is dominated by Lactobacillus spp., but a diverse array of other bacteria can be present in lower amounts. The activity of lactobacilli is essential to protect women from genital infections and to maintain the natural healthy balance of the vaginal micro ...

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Protocol for the Detection and Genotyping of Human Papillomaviruses Using a Liquid Bead Microarray Assay

More than 100 human papillomavirus (HPV) types have been identified, and over 40 of them infect the anogenital epithelium. Because each HPV type is associated with different risks for the development of cervical cancer, detecting and genotyping HPVs has increasingly become an integral pa ...

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Protocol for the Rapid Detection of the Urogenital Tract Mollicutes and Chlamydia with Concomitant LGV-(sub)typing

Urogenital tract infections can be caused by a number of pathogens, some of which, like the obligate intracellular Chlamydia trachomatis, are difficult to culture, or the cell wall-less mollicutes, like M. hominis or Ureaplasma spp. Real-time PCR (qPCR) has become an important diagnostic to ...

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Protocol for the Use of Enzyme-Linked Hybridization Assays for Genital Ulcer Disease

Etiologic agents of genital ulcer disease include herpes simplex 1 and 2, Treponema pallidum pallidum, Haemophilus ducreyi, and Klebsiella granulomatis. The advent of PCR has allowed for more rapid and sensitive detection of microbial pathogens. In this protocol, we describe the simul ...

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Protocol for the Use of a Rapid Real-Time PCR Method for the Detection of HIV-1 Proviral DNA Using Double-Stranded Primer

This chapter describes a real-time PCR method for the detection of HIV-1 proviral DNA in whole blood samples using a novel double-stranded primer system. The assay utilizes a simple commercially available DNA extraction method and a rapid and easy-to-perform real-time PCR protocol to cons ...

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Protocol for the Use of Self-Reporting Duplex Mutation Primers to Detect PCR Products in the Diagnosis of HBV

Quantitative measurements of serum hepatitis B virus (HBV) DNA are useful for tailoring of treatment schedules and the monitoring of HBV replication during therapy. We developed a novel fluorescence-based quantitative real-time PCR for quantitating HBV DNA based on the duplex mutat ...

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Protocol for the Use of Light Upon Extension Real-Time PCR for the Determination of Viral Load in HBV Infection

Real-time PCR has engendered wide acceptance for quantitation of hepatitis B virus (HBV) DNA in the blood due to its improved rapidity, sensitivity, reproducibility, and reduced contamination. Here we describe a cost-effective and highly sensitive HBV real-time quantitative assay b ...

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Protocol for the Use of a Silica Nanoparticle-Enhanced Microcantilever Sensor-Based Method to Detect HBV at Femtomolar Concentrations

DNA sensors that are capable of detecting specific DNA sequences in a bio-sample have recently been highlighted as a powerful and sensitive approach to detect infectious diseases caused by pathogens such as viruses and bacteria. Generally, DNA samples extracted from biological fluids ...

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Protocol for the Detection of Treponema pallidum in Paraffin-Embedded Specimens

Formalin-fixed paraffin-embedded (FFPE) tissue blocks are routinely used for histopathological examination and are also useful for specific pathogen detection by polymerase chain reaction (PCR). FFPE tissue is stable at ambient temperature for an extended period of time and rel ...

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Overview of Molecular Biological Methods for the Detection of Pathogens Causing Sexually Transmitted Infections

We review here different state-of-the-art molecular methods currently used in the diagnosis of sexually transmitted infections.

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Guidelines for the Use of Molecular Biological Methods to Detect Sexually Transmitted Pathogens in Cases of Suspected Sexual Abuse in Children

Testing for sexually transmitted infections (STIs) in children presents a number of problems for the practitioner that are not usually faced when testing adults for the same infections. The identification of an STI in a child, in addition to medical implications, can have serious legal impli ...

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Protocol for the Molecular Detection of Antibiotic Resistance Mechanisms in Neisseria gonorrhoeae

Gonorrhoea is no longer an easily treatable ailment but rather is now a challenging disease in terms of antimicrobial resistance (AMR) with treatment options rapidly diminishing. The causative agent of gonorrhoea, Neisseria gonorrhoeae, has managed to develop resistance to almost e ...

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