Simple monitoring of antiretroviral therapy with a signal-amplification-boosted HIV-1 p24 antigen assay with heat-denatured plasma

Böni, Jürg; Opravil, Milos; Tomasik, Zuzana; Rothen, Madeleine; Bisset, Leslie R.; Grob, Peter J.; Lüthy, Ruedi; Schüpbach, Jörg

doi:10.1097/00002030-199706000-00001

Cited by 74 publications

(43 citation statements)

References 21 publications

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“…A significant correlation between HD p24 Ag and HIV-1 RNA has been reported (3,19,23,28), primarily in studies evaluating HIV-1 subtype B. However, in several other studies the correlation was weak or absent (2,5,22).…”

Section: Discussionmentioning

confidence: 98%

“…The quantification of p24 antigen (Ag) has been considered for viral load monitoring (3,4,13,15,19,(21)(22)(23)(25)(26)(27)(28). The p24 Ag assay measures the virus directly and has been shown to correlate with levels of plasma HIV-1 RNA in untreated subjects (21,23,30) and persons receiving antiretroviral therapy (3,23,27,28).…”

mentioning

confidence: 99%

“…The p24 Ag assay measures the virus directly and has been shown to correlate with levels of plasma HIV-1 RNA in untreated subjects (21,23,30) and persons receiving antiretroviral therapy (3,23,27,28). In addition, detection of p24 Ag has been explored for early diagnosis of pediatric HIV-1 infection (19,20,24).…”

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confidence: 99%

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Detection and Quantification of Human Immunodeficiency Virus Type 1 p24 Antigen in Dried Whole Blood and Plasma on Filter Paper Stored under Various Conditions

Seidel

Coombs

et al. 2005

J Clin Microbiol

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The quantification of human immunodeficiency virus type 1 (HIV-1) by an assay measuring heat-dissociated (HD) p24 antigen (Ag) in specimens of whole blood and plasma stored on filter paper, and of plasma stored in tubes, was compared to HIV-1 RNA plasma levels determined by real-time reverse transcription (RT)-PCR. The stability of p24 Ag on filter paper under conditions simulating specimen transport was also evaluated. The HD p24 Ag in both plasma and whole-blood specimens stored on filter paper correlated with plasma HIV-1 RNA levels (Spearman rank ‫؍‬ 0.74 [P < 0.0001] and ‫؍‬ 0.56 [P ‫؍‬ 0.0001], respectively). The sensitivity of the HD p24 Ag assay was similar when plasma and whole blood on filter paper were contrasted to the real-time RT-PCR assay (80% versus 82.5% and 78.6% versus 83.3%, respectively). However, while the specificity of the HD p24 Ag assay of plasma on filter paper was 100%, the specificity was diminished in whole-blood specimens. The storage of specimens on filter paper for 2 weeks at 37°C, 24°C, or 0°C did not alter the detection or quantification of HD p24 Ag. These results suggest that transport and storage of plasma on filter paper and quantification of HD p24 Ag may be a reliable method for HIV-1 load monitoring.

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Section: Discussionmentioning

confidence: 98%

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confidence: 99%

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Detection and Quantification of Human Immunodeficiency Virus Type 1 p24 Antigen in Dried Whole Blood and Plasma on Filter Paper Stored under Various Conditions

Seidel

Coombs

et al. 2005

J Clin Microbiol

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“…The p24 antigen quantitation assay now includes heat dissociation of the interfering antigen-antibody immune complexes and subsequent signal amplification with biotyltyramide and streptavidin-horseradish peroxidase (S-HRP) to improve the sensitivity. The linear range of this assay has improved substantially with the introduction of a quantitative kinetic enzyme-linked immunosorbent assay (ELISA) reader and software (2,14,17,18,19,20,21). The ExaVir Load assay measures the activity of the virus-encoded enzyme RT, which is packaged together with the viral RNA in the HIV particle.…”

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confidence: 99%

Evaluation of Two Commercially Available, Inexpensive Alternative Assays Used for Assessing Viral Load in a Cohort of Human Immunodeficiency Virus Type 1 Subtype C-Infected Patients from South Africa

et al. 2005

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Although human immunodeficiency virus type 1 (HIV-1) RNA is the acknowledged "gold standard" marker for monitoring disease activity in patients receiving highly active antiretroviral therapy (HAART), it remains unaffordable in resource-constrained settings. The present study investigated two commercially available kits for the detection of HIV-1 viral load markers as more affordable alternatives to HIV-1 RNA quantitation. The greatly improved heat-denatured, signal-boosted HiSens HIV-1 p24 Ag Ultra kit (Perkin-Elmer) and the ExaVir Load Quantitative HIV-RT kit (Cavidi Tech AB) were compared with the Amplicor HIV-1 Monitor (version 1.5) assay (Roche Molecular Systems Inc.). A total of 117 samples containing HIV-1 subtype C were analyzed by all three methodologies. Eighty-nine of these samples represented serial measurements from 20 patients receiving HAART. The remaining samples analyzed were from a group of treatment-naïve patients. The association between the p24 antigen assay and the RNA assay was fairly strong (R 2 ‫؍‬ 0.686). The association between the reverse transcriptase (RT) quantitation assay and the RNA assay was strong (R 2 ‫؍‬ 0.810). Both alternative assays seemed most useful for the serial monitoring of patients receiving HAART (n ‫؍‬ 89 plasma samples from 20 patients), as all assays showed a statistically significant downward trend over time, with the trend being either linear or curvilinear. In addition, all three assays showed negative correlations with the CD4 count (CD4 count versus RNA load, r ‫؍‬ ؊0.336 and P ‫؍‬ 0.001; CD4 count versus p24 antigen level, r ‫؍‬ ؊0.541 and P < 0.0001; CD4 count versus RT level, r ‫؍‬ ؊0.358 and P ‫؍‬ 0.0006). Still of major concern are both the lack of sensitivity and the wide degrees of variability of both assays. However, both assays provide a less expensive alternative to the Roche viral load assay and demonstrate the same trends during treatment.

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“…However, HIV treatment budgets are under considerable pressure. Therefore, a sensitive immunoassay (Boni et al, 1997) for detection of the capsid protein (Von Sydow et al, 1988 ;Graziosi et al, 1993 ;Baumberger et al, 1993) would be an attractive and cheap alternative to more expensive molecular diagnostics. When resistance occurs, demonstrated by a re-activation of the virus, treatment can be changed to another combination of drugs.…”

Section: Introductionmentioning

confidence: 99%

Selection of human anti-human immunodeficiency virus type 1 envelope single-chain antibodies from a peripheral blood cell-based phage repertoire.

Haard

Kazemier

Oudshoorn³

et al. 1998

Journal of General Virology

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Monoclonal antibodies play an important role in the development of diagnostic assays. Instead of using hybridoma technology to isolate human immunodeficiency virus type 1-specific antibodies, a phagedisplayed antibody library was generated from a small number (10 7 ) of peripheral blood lymphocytes from a seropositive donor. Two families of singlechain antibodies (scFvs) were selected by biopanning with the envelope precursor gp160. ELISA and competition in the BlAcore system revealed that one antibody family recognized a conformationsensitive epitope within gp120, while the other antibody family was gp41-specific. The latter group had sequence similarity to antibodies recognizing

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Simple monitoring of antiretroviral therapy with a signal-amplification-boosted HIV-1 p24 antigen assay with heat-denatured plasma

Abstract: The performance of this antigen detection procedure is comparable to RNA PCR, thus providing a simple, high throughput alternative in monitoring the efficacy of antiretroviral treatment.

Cited by 74 publications

References 21 publications

Detection and Quantification of Human Immunodeficiency Virus Type 1 p24 Antigen in Dried Whole Blood and Plasma on Filter Paper Stored under Various Conditions

Detection and Quantification of Human Immunodeficiency Virus Type 1 p24 Antigen in Dried Whole Blood and Plasma on Filter Paper Stored under Various Conditions

Evaluation of Two Commercially Available, Inexpensive Alternative Assays Used for Assessing Viral Load in a Cohort of Human Immunodeficiency Virus Type 1 Subtype C-Infected Patients from South Africa

Selection of human anti-human immunodeficiency virus type 1 envelope single-chain antibodies from a peripheral blood cell-based phage repertoire.

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