A genotypic HIV-1 proviral DNA coreceptor tropism assay: Characterization in viremic subjects

Jennifer Brown, Harold Burger, Barbara Weiser, Richard B Pollard, Xiao Dong Li, Lynell J. Clancy, Russell E. Baumann, Amy A. Rogers, Hasnah B. Hamdan, Rick L. Pesano, Ron M. Kagan

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Background: HIV-1 coreceptor tropism testing is used to evaluate eligibility for CCR5 antagonist therapy. However, HIV-1 RNA-based tests are not suitable for virologically suppressed patients, therefore the use of proviral DNA tropism testing has been investigated. We describe a novel proviral DNA-based genotypic tropism assay and compare its performance to that of a sensitive HIV-1 RNA-based genotypic test. Methods: Tropism was determined using HIV-1 plasma RNA and proviral DNA from 42 paired samples from patients with plasma viral loads ≥1000 HIV-1 RNA copies/mL. Proviral DNA sample types included whole blood, separated peripheral blood mononuclear cells resuspended in phosphate-buffered saline and peripheral blood mononuclear cells resuspended in spun plasma. The HIV-1 envelope V3 region was PCR-amplified, sequenced in triplicate, and analyzed for tropism with the geno2pheno algorithm using a 10% false-positive rate (FPR). Results: Amplicons were obtained from proviral DNA and plasma RNA in 41/42 samples. Tropism predictions were highly concordant (93%-98%) between proviral DNA and plasma RNA, regardless of the proviral DNA isolation method. Non-R5 proviral DNA results were obtained for 100% of patients with detectable non-R5 plasma HIV-1 RNA results. Geno2pheno FPRs for proviral DNA and plasma RNA were highly correlated (Spearman rho = 0.86). Conclusions: Our findings demonstrate that proviral DNA tropism determinations from whole blood or peripheral blood mononuclear cells were highly concordant with plasma HIV-1 RNA tropism determinations. This assay may be useful for screening virologically suppressed patients for CCR5-antagonist eligibility and for research purposes.

Original languageEnglish (US)
Article number14
JournalAIDS Research and Therapy
Issue number1
StatePublished - May 21 2014


  • HIV-1 diagnostic tests
  • HIV-1 proviral tropism
  • HIV-1 tropism

ASJC Scopus subject areas

  • Virology
  • Molecular Medicine
  • Pharmacology (medical)


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