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Assessing the Suitability of Next-Generation Viral Outgrowth Assays to Measure Human Immunodeficiency Virus 1 Latent Reservoir Size.

  • Author(s): Stone, Mars;
  • Rosenbloom, Daniel IS;
  • Bacchetti, Peter;
  • Deng, Xutao;
  • Dimapasoc, Melanie;
  • Keating, Sheila;
  • Bakkour, Sonia;
  • Richman, Douglas D;
  • Mellors, John W;
  • Deeks, Steven G;
  • Lai, Jun;
  • Beg, Subul;
  • Siliciano, Janet D;
  • Pagliuzza, Amélie;
  • Chomont, Nicolas;
  • Lackman-Smith, Carol;
  • Ptak, Roger G;
  • Busch, Michael P
  • et al.

Published Web Location

https://academic.oup.com/jid/advance-article/doi/10.1093/infdis/jiaa089/5799268
No data is associated with this publication.
Abstract

Background

Evaluations of human immunodeficiency virus (HIV) curative interventions require reliable and efficient quantification of replication-competent latent reservoirs. The "classic" quantitative viral outgrowth assay (QVOA) has been regarded as the reference standard, although prohibitively resource and labor intensive. We compared 6 "next-generation" viral outgrowth assays, using polymerase chain reaction or ultrasensitive p24 to assess their suitability as scalable proxies for QVOA.

Methods

Next-generation QVOAs were compared with classic QVOA using single leukapheresis-derived samples from 5 antiretroviral therapy-suppressed HIV-infected participants and 1 HIV-uninfected control; each laboratory tested blinded batches of 3 frozen and 1 fresh sample. Markov chain Monte Carlo methods estimated extra-Poisson variation at aliquot, batch, and laboratory levels. Models also estimated the effect of testing frozen versus fresh samples.

Results

Next-generation QVOAs had similar estimates of variation to QVOA. Assays with ultrasensitive readout reported higher infectious units per million values than classic QVOA. Within-batch testing had 2.5-fold extra-Poisson variation (95% credible interval [CI], 2.1-3.5-fold) for next-generation assays. Between-laboratory variation increased extra-Poisson variation to 3.4-fold (95% CI, 2.6-5.4-fold). Frozen storage did not substantially alter infectious units per million values (-18%; 95% CI, -52% to 39%).

Conclusions

The data offer cautious support for use of next-generation QVOAs as proxies for more laborious QVOA, while providing greater sensitivities and dynamic ranges. Measurement of latent reservoirs in eradication strategies would benefit from high throughput and scalable assays.

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