Clinical validation of a quantitative HIV-1 DNA droplet digital PCR assay: Applications for detecting occult HIV-1 infection and monitoring cell-associated HIV-1 dynamics across different subtypes in HIV-1 prevention and cure trials

Powell, Laura; Dhummakupt, Adit; Siems, Lilly; Singh, Dolly; Duff, Yann Le; Uprety, Priyanka; Jennings, Cheryl; Szewczyk, Joseph; Chen, Ya; Nastouli, Eleni; Persaud, Deborah

doi:10.1016/j.jcv.2021.104822

Cited by 13 publications

(6 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is possible to collect and extract more significant volumes of blood to multiply the qPCR tests to improve sensitivity and accuracy, but this approach has limits in the case of the HIV reservoir explored in CD4 subpopulations and rare cells. Our results are in line with previous studies using dPCR for HIV DNA quantification 33 – 35 . dPCR techniques based on multiple targets could also further increase the accuracy of HIV DNA quantification 36 – 38 .…”

Section: Discussionsupporting

confidence: 93%

Accuracy of real-time PCR and digital PCR for the monitoring of total HIV DNA under prolonged antiretroviral therapy

Renault

Bolloré

Pisoni

et al. 2022

Sci Rep

View full text Add to dashboard Cite

Total HIV DNA is a standard marker to monitor the HIV reservoir in people living with HIV. We investigated HIV DNA quantification accuracy by a real-time PCR kit (qPCR) and digital PCR (dPCR) method within the same set of primers and probes. Among 48 aviremic patients followed for up to 7 years with qPCR, the mean coefficient of variation of total HIV DNA between two successive measurements was 77% (± 0.42log10 HIVDNA copies/106 PBMC). The total HIV DNA quantified by the two PCR methods has a high correlation (0.99 and 0.83, for 8E5 and PLHIV samples, respectively), but we observed better repeatability and reproducibility of the dPCR compared to the qPCR (CV of 11.9% vs. 24.7% for qPCR, p-value = 0.024). Furthermore, we highlighted a decay of the number of HIV copies in the 8E5 cell line qPCR standard over time (from 0.73 to 0.43 copies per cell), contributing to variations of HIV DNA results in patients whose HIV reservoir should be theoretically stabilized. Our study highlighted that absolute quantification of total HIV DNA by dPCR allows more accurate monitoring of the HIV reservoir than qPCR in patients under prolonged antiretroviral therapy.

show abstract

Section: Discussionsupporting

confidence: 93%

Accuracy of real-time PCR and digital PCR for the monitoring of total HIV DNA under prolonged antiretroviral therapy

Renault

Bolloré

Pisoni

et al. 2022

Sci Rep

View full text Add to dashboard Cite

show abstract

“…In IMPAACT P1115 clinical trial, neonates in Cohort 1 initiated very early ART at a median age of 7.8 h of life while neonates in Cohort 2 initiated very early ART at a median age of 32.8 h of life [30 ▪▪ ]. 64% (7/11) infants in Cohort 1 and 71% (5/7) infants in Cohort 2 of infants who maintained virologic control through 108 weeks of age reached undetectable HIV-1 DNA levels in PBMCs with an assay LOD of 0.6 log 10 cpm PBMCs [31]. In line with sex differences observed in Millar et al , in Cohort 1, female sex was associated with greater risk of virologic failure.…”

Section: Systematic Review Of Studies Measuring Total Hiv-1 Dna In Pe...supporting

confidence: 63%

“…After 48 weeks on ART, infants who initiated ART at <48 h and 49 h-14 days declined to a median 2.4 1og 10 cpm PBMC while those who initiated ART >14 days of life maintained HIV-1 DNA levels of 3.2 1og 10 cpm PBMC. This study found that smaller reservoir size in infants was associated with higher CD4 þ T-cell percentage and viral load <100 000 copies/ml pre-ART [ [31]. In line with sex differences observed in Millar et al, in Cohort 1, female sex was associated with greater risk of virologic failure.…”

Section: Key Pointssupporting

confidence: 58%

Age at ART initiation and proviral reservoir size in perinatal HIV-1 infection: considerations for ART-free remission

Bekka,

Kelly,

Haaren

et al. 2024

Current Opinion in HIV and AIDS

Self Cite

View full text Add to dashboard Cite

Purpose of review Achieving ART-free remission without the need for lifelong antiretroviral treatment (ART) is a new objective in HIV-1 therapeutics. This review comprehensively examines the literature to evaluate whether the age at ART initiation in children with perinatal HIV-1influences the size and decay of the HIV-1 reservoir. The insights gathered from this review serve to inform the field on the unique dynamics of HIV-1 reservoir size in perinatal HIV-1 infection as a function of age at ART initiation, as well as inform biomarker profiling and timing of ART-free remission strategies for children living with HIV-1 globally. Recent findings Recent studies demonstrate that initiating very early effective ART in neonates is feasible and limits HIV-1 reservoir size. The clinical relevance of limiting the HIV-1 reservoir size in perinatal infection was recently demonstrated in the Tatelo Study, which investigated a treatment switch from ART to two broadly neutralizing antibodies (bNAbs) in very early treated children. Low proviral reservoir size was associated with sustained virologic control for 24 weeks on bNAbs. Summary Immediate and early ART initiation for neonates and infants with perinatal HIV-1 is essential to restricting HIV-1 reservoir size that may enable ART-free remission.

show abstract

“…The expression levels of the target DNA in each droplet are determined by Poisson distribution thus eliminating the complications encountered in using standard curves providing improved sensitivity and precision over qPCR 22 . The assay has recently been clinically validated across different HIV-1 subtypes 34 . Additional refinements have used multiplexed dPCR in the Intact Proviral DNA assay (IPDA), to effectively discriminate intact from defective proviruses 22 , 35 .…”

Section: Introductionmentioning

confidence: 99%

Use of laboratory-developed assays in global HIV-1 treatment-monitoring and research

Malisa

Manak²,

Michelo³

et al. 2023

Sci Rep

View full text Add to dashboard Cite

There has been a surge in the emergence of HIV-1 drug resistance in Low and Middle-Income Countries (LMICs) due to poor drug-adherence and limited access to viral load testing, the current standard for treatment-monitoring. It is estimated that only 75% of people living with HIV (PLWH) worldwide have access to viral load testing. In LMICs, this figure is below 50%. In a recent WHO survey in mostly LMICs, 21 out of 30 countries surveyed found HIV-1 first-line pre-treatment drug resistance in over 10% of study participants. In the worst-affected regions, up to 68% of infants born to HIV-1 positive mothers were found to harbour first-line HIV-1 treatment resistance. This is a huge public health concern. Greater access to treatment-monitoring is required in LMICs if the UNAIDS “third 95” targets are to be achieved by 2030. Here, we review the current challenges of viral load testing and present the case for greater utilization of Laboratory-based assays that quantify intracellular HIV-1 RNA and/or DNA to provide broader worldwide access to HIV-1 surveillance, drug-resistance monitoring, and cure-research.

show abstract

Clinical validation of a quantitative HIV-1 DNA droplet digital PCR assay: Applications for detecting occult HIV-1 infection and monitoring cell-associated HIV-1 dynamics across different subtypes in HIV-1 prevention and cure trials

Cited by 13 publications

References 33 publications

Accuracy of real-time PCR and digital PCR for the monitoring of total HIV DNA under prolonged antiretroviral therapy

Accuracy of real-time PCR and digital PCR for the monitoring of total HIV DNA under prolonged antiretroviral therapy

Age at ART initiation and proviral reservoir size in perinatal HIV-1 infection: considerations for ART-free remission

Use of laboratory-developed assays in global HIV-1 treatment-monitoring and research

Contact Info

Product

Resources

About