Details

Autor(en) / Beteiligte

• Levy, Claire N.
• Hughes, Sean M.
• Roychoudhury, Pavitra
• Reeves, Daniel B.
• Amstuz, Chelsea
• Zhu, Haiying
• Huang, Meei-Li
• Wei, Yulun
• Bull, Marta E.
• Cassidy, Noah A.J.
• McClure, Jan
• Frenkel, Lisa M.
• Stone, Mars
• Bakkour, Sonia
• Wonderlich, Elizabeth R.
• Busch, Michael P.
• Deeks, Steven G.
• Schiffer, Joshua T.
• Coombs, Robert W.
• Lehman, Dara A.
• Jerome, Keith R.
• Hladik, Florian

Titel

A highly multiplexed droplet digital PCR assay to measure the intact HIV-1 proviral reservoir

Ist Teil von

• Cell reports. Medicine, 2021-04, Vol.2 (4), p.100243-100243, Article 100243

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2021

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• Quantifying the replication-competent HIV reservoir is essential for evaluating curative strategies. Viral outgrowth assays (VOAs) underestimate the reservoir because they fail to induce all replication-competent proviruses. Single- or double-region HIV DNA assays overestimate it because they fail to exclude many defective proviruses. We designed two triplex droplet digital PCR assays, each with 2 unique targets and 1 in common, and normalize the results to PCR-based T cell counts. Both HIV assays are specific, sensitive, and reproducible. Together, they estimate the number of proviruses containing all five primer-probe regions. Our 5-target results are on average 12.1-fold higher than and correlate with paired quantitative VOA (Spearman's ρ = 0.48) but estimate a markedly smaller reservoir than previous DNA assays. In patients on antiretroviral therapy, decay rates in blood CD4+ T cells are faster for intact than for defective proviruses, and intact provirus frequencies are similar in mucosal and circulating T cells. [Display omitted] HIV cure studies rely on precise HIV reservoir assays that can be done at scaleMultiplexed droplet digital PCR can probe multiple HIV genome targets per sampleHIV-1 proviruses containing 5 probed genomic regions are likely intactPCR-based counting of T cells improves reservoir quantification in tissues Evaluation of HIV cure interventions requires accurate and scalable measurement of the replication-competent HIV reservoir. Levy at al. describe a highly multiplexed droplet digital PCR assay that simultaneously quantifies likely intact HIV-1 proviruses and T lymphocytes. They validate the assay in cell and tissue specimens from several patient cohorts.