Clinical implications of elevated HIV-1 viral load results obtained from samples stored frozen in vacutainer plasma preparation tubes

Cloherty, Gavin; Swanson, Priscilla; Lucic, Danijela; Dieckhaus, Kevin D.; Anthony, Paul; Cataline, Philip R.; Herman, Christine; Hackett, John; Skolnik, Paul R.; Chirch, Lisa M.

doi:10.1016/j.jviromet.2014.01.025

Cited by 5 publications

(5 citation statements)

References 6 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1B and D ). Notably, the use of the Roche TaqMan assay did not overestimate the quantity of HIV-1 RNA present in the culture supernatants, as indicated by the cross-validation test using the Abbott RealTime HIV-1 assay, which avoids the potential detection of HIV-1 DNA ( 28 ) ( Fig. 3 ).…”

Section: Resultsmentioning

confidence: 95%

In Vitro Reactivation of Replication-Competent and Infectious HIV-1 by Histone Deacetylase Inhibitors

Banga

Procopio

Cavassini

et al. 2016

J Virol

View full text Add to dashboard Cite

The existence of long-lived HIV-1-infected resting memory CD4 T cells is thought to be the primary obstacle to HIV-1 eradication. In the search for novel therapeutic approaches that may reverse HIV-1 latency, inhibitors of histone deacetylases (HDACis) have been tested to reactivate HIV-1 replication with the objective of rendering HIV-1-infected cells susceptible to elimination either by HIV-specific CD8 T cells or through virus-mediated cytopathicity. In the present study, we evaluated the efficiency of HDACis to reactivate HIV-1 replication from resting memory CD4 T cells isolated from aviremic long-term-treated HIV-1-infected subjects. We demonstrate that following prolonged/repeated treatment of resting memory CD4 T cells with HDACis, HIV-1 replication may be induced from primary resting memory CD4 T cells isolated from aviremic long-term-treated HIV-1-infected subjects. More importantly, we demonstrate that HIV-1 reactivated in the cell cultures was not only replication competent but also infectious. Interestingly, givinostat, an HDACi that has not been investigated in clinical trials, was more efficient than vorinostat, panobinostat, and romidepsin in reversing HIV-1 latency in vitro. Taken together, these results support further evaluation of givinostat as a latency-reversing agent (LRA) in aviremic long-term-treated HIV-1-infected subjects.IMPORTANCE The major barrier to HIV cure is the existence of long-lived latently HIV-1-infected resting memory CD4 T cells. Latently HIV-1-infected CD4 T cells are transcriptionally silent and are therefore not targeted by conventional antiretroviral therapy (ART) or the immune system. In this context, one strategy to target latently infected cells is based on pharmacological molecules that may force the virus to replicate and would therefore render HIV-1-infected cells susceptible to elimination either by HIV-specific CD8 T cells or through virus-mediated cytopathicity. In this context, we developed an experimental strategy that would allow the evaluation of latency-reversing agent (LRA) efficiency in vitro using primary CD4 T cells. In the present study, we demonstrate that HDACis are potent inducers of replication-competent and infectious HIV-1 in resting memory CD4 T cells of long-term ART-treated patients and identify givinostat as the most efficient LRA tested.

show abstract

Section: Resultsmentioning

confidence: 95%

In Vitro Reactivation of Replication-Competent and Infectious HIV-1 by Histone Deacetylase Inhibitors

Banga

Procopio

Cavassini

et al. 2016

J Virol

View full text Add to dashboard Cite

show abstract

“…Four published studies evaluated PPTs on viral load assays that were commercially available at the time of this review (Abbott RealTi m e HIV-1 and Roche COBAS AmpliPREP/COBAS TaqMan HIV-1 Test, v2.0) [25–28]. The three studies evaluating the Abbott viral load assay showed no significant change in viral load results regardless of if the PPTs were frozen and thawed or transported after initial centrifugation and prior to testing [25–27]. The three studies assessing the Roche assay found elevated viral load results if the PPTs were frozen or transported without a second centrifugation prior to testing [26–28].…”

Section: Resultsmentioning

confidence: 99%

Systematic review of the accuracy of plasma preparation tubes for HIV viral load testing

et al. 2019

View full text Add to dashboard Cite

Expanding access to HIV viral load testing is essential to improving the care and treatment of people living with HIV/AIDS and ending the AIDS epidemic. Though significant investments have been made in the past five years, many high burden, low resource countries continue to have viral load access rates below 50%. Plasma preparation tubes (PPTs) can simplify storage, transport, and preparation of plasma used for viral load testing. A systematic review was conducted to evaluate the accuracy of plasma preparation tubes for HIV viral load testing. Study results regarding the accuracy of PPT viral load measurements across various storage and transportation conditions were examined. The quality of evidence was evaluated using GRADE and QUADAS-2 criteria. The review identified 16 studies using PPTs with data from 6,141 individuals from 1995 to 2014. Overall the quality of evidence was rated as moderate, with unclear applicability for studies evaluating viral load assays that are no longer commercially available. Significantly elevated viral load results (>0.3 log copies/ml difference) have been observed with PPTs; however, when manufacturer handling instructions are followed, when plasma is aliquoted into a secondary tube, or when PPTs are centrifuged prior to testing, PPT results only differed from standard EDTA plasma testing using commercially available viral load assays by a range on average of -0.03 to +0.08 log copies/ml across studies. Although spuriously elevated viral load results have been observed with PPTs, following proper sample handing techniques have been shown to provide accurate results. PPTs, therefore, provide a high quality alternative specimen type for countries seeking solutions to infrastructure and specimen transportation challenges in an effort to scale-up viral load testing and achieve 90-90-90 targets.

show abstract

“…However, these models are highly complex and currently available statistical software is not capable of handling these models. We also should acknowledge that some of these pVL assays may be susceptible to artefacts such as viral diversity, virus subtype, and primers used in primary plasma preparation tubes, which may contribute to the measurement variability here described [ 21 – 26 ]. Although these are legitimate concerns, it should be noted that the virology laboratory does not use these tubes, the same primers were used in all measurements, and >90% of our patients have clade B subtype.…”

Section: Discussionmentioning

confidence: 99%

Estimation of measurement error in plasma HIV-1 RNA assays near their limit of quantification

et al. 2017

View full text Add to dashboard Cite

BackgroundPlasma HIV-1 RNA levels (pVLs), routinely used for clinical management, are influenced by measurement error (ME) due to physiologic and assay variation.ObjectiveTo assess the ME of the COBAS HIV-1 Ampliprep AMPLICOR MONITOR ultrasensitive assay version 1.5 and the COBAS Ampliprep Taqman HIV-1 assay versions 1.0 and 2.0 close to their lower limit of detection. Secondly to examine whether there was any evidence that pVL measurements closest to the lower limit of quantification, where clinical decisions are made, were susceptible to a higher degree of random noise than the remaining range.MethodsWe analysed longitudinal pVL of treatment-naïve patients from British Columbia, Canada, during their first six months on treatment, for time periods when each assay was uniquely available: Period 1 (Amplicor): 08/03/2000–01/02/2008; Period 2 (Taqman v1.0): 07/01/2010–07/03/2012; Period 3 (Taqman v2.0): 08/03/2012–30/06/2014. ME was estimated via generalized additive mixed effects models, adjusting for several clinical and demographic variables and follow-up time.ResultsThe ME associated with each assay was approximately 0.5 log10 copies/mL. The number of pVL measurements, at a given pVL value, was not randomly distributed; values ≤250 copies/mL were strongly systematically overrepresented in all assays, with the prevalence decreasing monotonically as the pVL increased. Model residuals for pVL ≤250 copies/mL were approximately three times higher than that for the higher range, and pVL measurements in this range could not be modelled effectively due to considerable random noise of the data.ConclusionsAlthough the ME was stable across assays, there is substantial increase in random noise in measuring pVL close to the lower level of detection. These findings have important clinical significance, especially in the range where key clinical decisions are made. Thus, pVL values ≤250 copies/mL should not be taken as the “truth” and repeat pVL measurement is encouraged to confirm viral suppression.

show abstract

Clinical implications of elevated HIV-1 viral load results obtained from samples stored frozen in vacutainer plasma preparation tubes

Cited by 5 publications

References 6 publications

In Vitro Reactivation of Replication-Competent and Infectious HIV-1 by Histone Deacetylase Inhibitors

In Vitro Reactivation of Replication-Competent and Infectious HIV-1 by Histone Deacetylase Inhibitors

Systematic review of the accuracy of plasma preparation tubes for HIV viral load testing

Estimation of measurement error in plasma HIV-1 RNA assays near their limit of quantification

Contact Info

Product

Resources

About