The ARTμS: a novel microfluidic CD4+ T-cell enumeration system for monitoring antiretroviral therapy in HIV patients

Lab Chip. 2016 Feb 7;16(3):506-14. doi: 10.1039/c5lc01153a.

Abstract

We report on a novel and cost-effective microfluidic platform that integrates immunomagnetic separation and cell enumeration via DNA-induced bead aggregation. Using a two-stage immunocapture microdevice, 10 μL of whole blood was processed to isolate CD4+ T-cells. The first stage involved the immuno-subtraction of monocytes by anti-CD14 magnetic beads, followed by CD4+ T-cell capture with anti-CD4 magnetic beads. The super hydrophilic surface generated during polydimethylsiloxane (PDMS) plasma treatment allowed for accurate metering of the CD4+ T-cell lysate, which then interacted with silica-coated magnetic beads under chaotropic conditions to form aggregates. Images of the resulting aggregates were captured and processed to reveal the mass of DNA, which was used to back-calculate the CD4+ T-cell number. Studies with clinical samples revealed that the analysis of blood within 24 hours of phlebotomy yielded the best results. Under these conditions, an accurate cell count was achieved (R(2) = 0.98) when compared to cell enumeration via flow cytometry, and over a functional dynamic range from 106-2337 cells per μL.

MeSH terms

  • Anti-Retroviral Agents / administration & dosage*
  • CD4 Lymphocyte Count / instrumentation
  • CD4 Lymphocyte Count / methods
  • Female
  • HIV Infections* / drug therapy
  • HIV Infections* / immunology
  • Humans
  • Male
  • Microfluidic Analytical Techniques* / instrumentation
  • Microfluidic Analytical Techniques* / methods
  • Monitoring, Physiologic* / instrumentation
  • Monitoring, Physiologic* / methods

Substances

  • Anti-Retroviral Agents