The Infona portal uses cookies, i.e. strings of text saved by a browser on the user's device. The portal can access those files and use them to remember the user's data, such as their chosen settings (screen view, interface language, etc.), or their login data. By using the Infona portal the user accepts automatic saving and using this information for portal operation purposes. More information on the subject can be found in the Privacy Policy and Terms of Service. By closing this window the user confirms that they have read the information on cookie usage, and they accept the privacy policy and the way cookies are used by the portal. You can change the cookie settings in your browser.
A DNA heteroduplex tracking assay (HTA) using single-stranded probes is described. This assay provides a rapid means of resolving genetic variants coamplified by PCR and of measuring the level of particular variants in complex populations. To confidently detect minor quasispecies changes, the importance of maximizing template input into nested PCR (nPCR) and of duplicating nPCR and HTA to ensure correct...
Antigen capture enzyme-linked immunosorbent assay (ELISA) for quantitation of the p24 gag protein of human immunodeficiency virus type-1 (HIV-1) is currently the most common method used to demonstrate virus replication bothin vivoandin vitro.The present paper describes an ELISA employing readily available inexpensive reagents and gives detailed suggestions for optimizing the variables for specific...
We describe a method for the production of high-titer stocks of human immunodeficiency virus type 1 (HIV-1) pseudotyped with vesicular stomatitis virus envelope glycoprotein (VSV G). VSV G pseudotypes provide several advantages over other retroviral envelope proteins. The VSV G envelope is mechanically stable, enabling ultracentrifugal concentration of virions to high titers, and VSV G has a broad...
The SCID-hu mouse bearing a functional human thymic implant can be easily infected with HIV. Infection results in virus replication and relatively rapid depletion of CD4 + human thymocytes, resulting in a pathologic profile similar to that seen in the thymus of HIV-infected humans. The use of the SCID-hu model for HIV research requires protection of the animals from opportunistic infections...
A technique is described for detecting the activity of neutralizing polyclonal or monoclonal antibodies against HIV-1 primary isolates. Most commonly, neutralizing antibody activity for HIV-1 is assessed by quantifying the ability of antibodies to inhibit virus infection in mitogen-activated peripheral blood mononuclear cells or transformed lymphocytes. Because the target of HIV infectionin vivois...
Most catalytically active human immunodeficiency virus (HIV) reverse transcriptase (RT) mutants characterized to date have been isolated from the virus after treatment with HIV RT inhibitors such as nucleoside analogs. However, detailed understanding of structure–function relationships, and of the roles of the several catalytic activities of HIV RT in viral replication, requires characterization of...
Retroviruses evolve at rapid rates. This allows them to escape immune surveillance, thwarts vaccine development, and leads to rapid emergence of drug-resistant virus. Information regarding the retroviral mutation rates and the underlying mechanisms of mutagenesis will undoubtedly expedite the development of strategies to combat retroviral-mediated diseases. In this review, we discuss how the unique...
Integration of retroviral DNA, an essential step during the retroviral life cycle, is mediated by the viral protein integrase. Simplein vitroassays for measuring integrase activities are described, including catalysis (3′-end processing, 3′-end joining, disintegration), juxtaposition of viral DNA ends, DNA binding, and target site selection. The described assays will be useful in elucidating the molecular...
Quantitative competitive PCR is a highly sensitive technique that allows accurate quantitation of small amounts of RNA. We have modified the original method to include the use of an internal standard at all stages of sample analysis. In this way, the method can accommodate for variations in the recovery of viral particles and in the isolation of genomic RNA as well as provide a suitable competitive...
In vivochallenge procedures can be very useful in the analysis of allergic symptoms. Skin testing has a high degree of sensitivity and specificity for determining antigens that cause allergic disease. However, positive skin tests do not necessarily indicate that a specific allergen causes symptoms specific for a certain organ. Nasal and whole lung provocation testing can help define relevant allergens...
Environmental specimens (dust) from indoor home, school, and work-place environments can be evaluated for the content of aeroallergens produced by dust mite, cat, dog, cockroach, and molds, as a means of determining exposure risk and facilitating avoidance therapy. This article examines the variables that influence the levels of these allergens in indoor environments, methods for sampling, clinical...
Allergen extracts are prepared from a wide variety of source materials including pollens, fungi, arthropods, animal danders, foods, and dusts. The composition of allergen extracts can vary depending on the allergen source, manufacturing process, and storage conditions. Allergen-specific immunoglobulin E (IgE) assays and skin tests employ a variety of allergen-containing reagents that confer specificity...
Mast cells are the primary effector cells of immediate hypersensitivity reactions in humans. Upon mast cell activation both preformed and newly synthesized mediators are secreted. Histamine can be measured by fluorometric assays, radioenzymatic assays, and immunoassays. These methods have been applied to plasma and urine to detect histamine that had been releasedin vivoand to release histaminein vitrofrom...
As with all diagnostic laboratory testing, some form of external proficiency testing is required both for the laboratory undertaking the testing and for the safety of the requesting clinician and his patient. Allergen Specific immunoglobulin E antibody testing in serum can produce different results depending on the method. Originally External Quality Assessment schemes in Europe were run on a National...
Products derived from eosinophils, basophils, and mast cells are considered critical to the development of allergic diseases. Studies of the selective recruitment, accumulation, and/or activation of these cells during human allergic inflammatory reactionsin vitroandin vivohave been facilitated by a wide variety of methods. Some have been developed to identify and isolate these cells from a variety...
The clinical immunology laboratory provides support to the allergist in the diagnosis and management of human allergic diseases. Following a clinical history, the detection of allergen-specific immunoglobulin E (IgE) in serum can be useful in the definitive diagnosis of an IgE antibody-mediated hypersensitivity. Total serum IgE, the multiallergen screen, and mast cell tryptase are less commonly measured...
The pathophysiology of allergic disease is multifactorial, involving an intricate network of interactions among cells, mediators, and cytokines. Substantial progress has been made in defining the role of antigen-specific T cells and cytokines in the regulation of immunoglobulin E (IgE) synthesis and the atopic diseases. The development of antigen-specific T-cell lines and clones has facilitated efforts...
Gene targeting, which permits alteration of a chosen gene in a predetermined way by homologous recombination, is an emerging technology in malaria research. Soon after the development of techniques for stable transformation of red blood cell stages ofPlasmodium falciparumandPlasmodium berghei,genes of interest were disrupted in the two species. The main limitations of gene targeting in malaria parasites...
Set the date range to filter the displayed results. You can set a starting date, ending date or both. You can enter the dates manually or choose them from the calendar.