Cutaneous Manifestations of Sexually Transmitted Disease in the HIV-Positive Patient
Clay J. Cockerell, Antoanella Calame in Cutaneous Manifestations of HIV Disease, 2012
While still uncommon, since 2003 LGV has been reported with increased frequency in the industrialized world in certain populations such as MSM, particularly those practicing receptive anal intercourse.97–102 Retrospective studies on anal swabs taken from STD clinic visitors in San Francisco and Amsterdam revealed that the current epidemic is caused by the L2b strain of C. trachomatis.103,104 Presence of the L2b has been documented since 1981 in the US and since 2000 in Europe, leading some in the field to declare this a ‘slow epidemic’.105 The undetected spread of LGV among the MSM population is attributed to testing procedures employed prior to 2003, when positive anal swabs were recorded simply as Chlamydia proctitis without the performance of further serotyping. This was due to the presumed paucity of cases of LGV outside of endemic countries.105 Recently developed real-time polymerase chain reaction (RT-PCR) test designed specifically for LGV serotypes can lessen diagnostic difficulties as the test is rapid, highly specific, and can be performed under routine microbiological laboratory conditions.106
Water monitoring and testing *
Jamie Bartram, Rachel Baum, Peter A. Coclanis, David M. Gute, David Kay, Stéphanie McFadyen, Katherine Pond, William Robertson, Michael J. Rouse in Routledge Handbook of Water and Health, 2015
Increasingly it has been recognized that there is no direct correlation between numbers of any indicator and enteric pathogens. As a consequence of the possible ambiguity of the term ‘microbial indicator’, Ashbolt et al. (2001) note that the following terms are now recognized: 1. general (process) microbial indicators, 2. fecal indicators and 3. index and model organisms. Meanwhile, recent advances in molecular microbiology have supported the standardization and automation of methods to enumerate genes copies from specific waterborne pathogens (Girones et al., 2010) and tools are emerging that may further elucidate the ecological behavior of enteric organisms beyond the gut environment. Although concerns continue to be expressed about the uncertain relationship between numbers of infectious organisms and concentrations of nucleic acid targets in environmental samples, quantitative real-time polymerase chain reaction (qPCR) is now extensively used to support the burgeoning field of quantitative microbial risk assessment (QMRA). In addition, in the evolving field of quantitative microbial source tracking (QMST) the identification and enumeration of microbial species and groups of specifically human origin (or other specified animal groups) have begun to inform decision-making in the fields of watershed and recreational water management.
Understanding and Diagnosing Herpes Simplex Virus
Marie Studahl, Paola Cinque, Tomas Bergström in Herpes Simplex Viruses, 2017
A revolution has taken place in laboratory techniques for herpes simplex virus (HSV) diagnosis in recent years (1–3), and it is now possible to produce accurate results within a day, sometimes within hours (4,5). This development is not only driven by the increase in the number of immunosuppressed patients, in whom HSV infections are responsible for significant morbidity and mortality (6–10), but also by the introduction of safe antiviral drugs for the treatment of both acute and chronic HSV infections (11–14). These factors have contributed to a change in the clinician’s expectations of diagnostic virology and have created a welcome opportunity for the medical virologist to develop rapid, accurate, and relevant test modalities. Examples of these are nucleic acid (NA) amplification techniques, including HSV DNA quantitative assays (11,15–17), and the introduction of sophisticated laboratory services such as antiviral susceptibility testing (18,19). These improved laboratory services are rapidly redefining the natural history and epidemiological picture of HSV infection (20–24). As clinicians become increasingly reliant on these techniques, the clinical need for robust molecular techniques, such as fully automated real-time polymerase chain reaction (PCR) assays (25) will become evident.
Multiomic analysis of cytokines in immuno-oncology
Published in Expert Review of Proteomics, 2020
Real-time polymerase chain reaction (PCR) is a widely used method in molecular biology that began in the early 1980s. It is used to detect nucleic acid sequences using DNA polymerases and primers. In this way it can detect individual nucleotide mutations and can confirm gene expression [121,122]. Based on genetic analyses using real-time reverse transcription polymerase chain reaction (RT-PCR), the quantification of gene products can be estimated [123]. This is a widely used method for testing cytokines in tissues, cell lysates, cell cultures, and from biopsy materials obtained from patients. Most researchers use this method to compare the relative differences in cytokines between samples or more frequently to compare the obtained values of a gene transcript for cytokines after in vitro cell stimulation [45]. Hovewer, this method can also be used as a quantitative measure because the amount of fluorescence is directly proportional to the amount of product generated in the PCR cycle.
Pre-imaginal exposure to mancozeb induces morphological and behavioral deficits and oxidative damage in Drosophila melanogaster
Published in Drug and Chemical Toxicology, 2023
Cynthia Camila Ziech, Nathane Rosa Rodrigues, Giulianna Echeverria Macedo, Karen Kich Gomes, Illana Kemmerich Martins, Jeferson Luis Franco, Thaís Posser
Expression of Nrf2, NFkB, p38β, Hsp83, Akt1, Gpdh, dIlP2, dIlP6, FOXO, tor, per, and tim genes of newly emerged Drosophila were analyzed (Table 1). Trizol Reagent (Invitrogen™) was used to extract about 1 µg of total RNA from 20 flies, according to the manufacturer’s protocol, and then treated with DNAse I (DNAse I Amplification Grade – Invitrogen™, NY). cDNA was synthesized with iScript cDNA Synthesis Kit and random primers again according to the manufacturer’s protocol (Biorad© CA, USA). The quantitative real-time polymerase chain reaction was performed according to the protocol previously described by Rodrigues et al. (2018), with the exception of the tubulin gene, used as an endogenous reference gene in this study, presenting no alterations in response to the treatment.
Estrogen receptor gene polymorphisms and their influence on clinical status of Caucasian patients with primary open angle glaucoma
Published in Ophthalmic Genetics, 2019
Ewa Kosior-Jarecka, Małgorzata Sagan, Dominika Wróbel-Dudzińska, Urszula Łukasik, Tin Aung, Chiea Chuen Khor, Janusz Kocki, Tomasz Żarnowski
DNA was isolated from peripheral blood by means of QIAamp DNA Blood Midi Kit (QIAGEN Inc., Germany). DNA concentration was measured using NanoDrop 2000/2000c Spectrophotometer V1.0 (Thermo Fisher Scientific). TaqMan SNP probe (Applied Biosystems) and CFX96 Real-Time PCR Detection System thermal cycler (Bio-Rad) were used to assay 2 polymorphisms of the single nucleotide of the ESR1 gene and ESR2 polymorphisms of single nucleotide of the ESR2 gene. DNA amplification was obtained by means of a Real-Time Polymerase Chain Reaction. Real-Time PCR Thermal Cycler is equipped with optical system, which tracks PCR process in real time. To this end, fluorochrome-labeled molecular TaqMan probes (oligonucleotides of about 20–30 bp length) complementary to replicated DNA sequences and to PCR product were used. At the end of 5′ probe there is a fluorescent dye and at the end of 3′ a quencher. The dyes that were used in the study were FAM (6-carboxyfluorescein) and VIC (ABI company trademark, USA) and the quencher was TAMRA (6-carboxytetramethylrhodamine). Probes and primers were designed by Applied Biosystems (Foster City, CA, USA).
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