A rapid and simple method for extracting human immunodeficiency virus type 1 RNA from plasma: enhanced sensitivity

Mulder, John E.; Resnick, Robert H.; Saget, B; Scheibel, Steven F.; Herman, Steven A.; Payne, H. Ross; Harrigan, Richard; Kwok, Shirley

doi:10.1128/jcm.35.5.1278-1280.1997

Cited by 75 publications

(27 citation statements)

References 7 publications

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“…This procedure has an analytical sensitivity for detection of HIV-1 RNA in plasma at 20 copies per mL. 15 Six individuals (four men and two women) were selected. The mean age at the discovery of HIV infection was 27 years (range, 21-32 years).…”

Section: Methodsmentioning

confidence: 99%

HIV antibody screening remains indispensable for ensuring viral safety of blood components despite NAT implementation

et al. 2003

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Section: Methodsmentioning

confidence: 99%

HIV antibody screening remains indispensable for ensuring viral safety of blood components despite NAT implementation

et al. 2003

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“…The RNA was precipitated with 1 ml of isopropanol at room temperature at 14,000 g for 30 min, washed with 70 % ethanol, and resuspended in 100 p1 diethylpyrocarbonate-treated H20. Samples were stored on ice until amplification [18]. Of each sample, 5 pl was used for cDNA generation with AMV-RT (Boehringer Mannheim, Germany), as described by the manufacturer.…”

Section: Postoperative Monitoringmentioning

confidence: 99%

Analysis of potential porcine endogenous retrovirus (PERV) transmission in a whole-organ xenotransplantation model without interfering microchimerism

et al. 2001

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The question whether porcine xenografts can lead to porcine endogenous retrovirus (PERV) infection of recipients is critical for the evaluation of the safety of pigto-man xenotransplantation. Unfortunately, polymerase chain reaction (PCR)-based analysis of potential PERV infections in nonhuman-primate whole-organ xenotransplantation models is hampered by false positive results due to chimeric porcine cells. To avoid the inherent analytical problem of xenomicrochimerism, we developed a non-life-supporting pig-to-primate kidney xenotransplantation model: porcine kidneys were transplanted, whereas the functioning recipient kidneys remained in situ. Subsequent to rejection (after 2 hours to 15 days), xenografts were removed, and recipients remained alive for up to 287 days. Immunosuppressive therapy based on cyclophosphamide, cyclosporine, and steroids was maintained for 28 days after transplantation. Using appropriate PCR assays, xenochimerism was found in tissue samples and partly even in peripheral blood leukocytes (PBLs) while the porcine kidneys were in situ. After graft removal, xenochimerism was no longer detectable, thus allowing analysis for possible PERV transmission.

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“…Virus load was quantified from cryopreserved plasma using the Amplicor RT-PCR kit (Roche Diagnostic Systems, USA, detection limit of 50 copies/ml) for patients SC19 and S1 and using standard Amplicor HIV-1 monitor test, version 1.5 (Roche, Rotkreuz, Switzerland) with an ultrasensitive modification [29,30] (detection limit of 20 copies/ml) for patient Z18.…”

Section: Determination Of Virus Loadmentioning

confidence: 99%

Directex vivo analysis reveals distinct phenotypic patterns of HIV-specific CD8+ T lymphocyte activation in response to therapeutic manipulation of virus load

et al. 2001

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Therapeutic intervention with antiretroviral therapy (ART) enables the modulation of HIV virus load and hence provides a unique opportunity to study the consequences of varying antigen load onthe phenotype of virus‐specific CD8+ T lymphocytes in a persistent human viral infection. The recent advent of tetrameric peptide / HLA class I complexes has enabled the direct phenotypic characterization of antigen‐specific T cell populations ex vivo. Here, we use this technology to examine directly ex vivo the consequences of therapeutic manipulation of HIV virus load on the phenotype of HIV‐specific CTL. Our observations show that: (1) distinct sequential activation patterns of CD8+ T cells are associated with increasing virus load; (2) T cell receptor (TCR) down‐regulation without apoptosis represents an early event during the generation of a T cell response in a natural infection and precedes the emergence of two distinct antigen‐specific CD8+ T cell populations which differ in TCR and CD8 expression levels. Clear differences in surface Annexin V staining were observed between these populations. The observation that CTL activation, demonstrated by TCR and CD8 down‐regulation, in response to rising levels of virus load, co‐segregates with apoptosis only during later stages of the response indicates that antigen‐associated cell death is restricted to distinct subpopulations of CTL.

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A rapid and simple method for extracting human immunodeficiency virus type 1 RNA from plasma: enhanced sensitivity

Cited by 75 publications

References 7 publications

HIV antibody screening remains indispensable for ensuring viral safety of blood components despite NAT implementation

HIV antibody screening remains indispensable for ensuring viral safety of blood components despite NAT implementation

Analysis of potential porcine endogenous retrovirus (PERV) transmission in a whole-organ xenotransplantation model without interfering microchimerism

Directex vivo analysis reveals distinct phenotypic patterns of HIV-specific CD8+ T lymphocyte activation in response to therapeutic manipulation of virus load

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