Evaluation of dynabeads and cytospheres compared with flow cytometry to enumerate CD4+ T cells in HIV-infected ugandans on antiretroviral therapy

Fred Lutwama, Ronnie Serwadda, Harriet Mayanja-Kizza, Hasan M. Shihab, Allan Ronald, Moses R. Kamya, David Thomas, Elizabeth Johnson, Thomas C. Quinn, Richard D. Moore, Lisa A. Spacek

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Laboratory-based monitoring of antiretroviral therapy is essential but adds a significant cost to HIV care. The World Health Organization 2006 guidelines support the use of CD4 lymphocyte count (CD4) to define treatment failure in resource-limited settings. METHODS: We compared CD4 obtained on replicate samples from 497 HIV-positive Ugandans (before and during ART) followed for 18 months by 2 manual bead-based assays, Dynabeads (Dynal Biotech), and Cytospheres (Beckman Coulter) with those generated by flow cytometry at the Infectious Diseases Institute in Kampala, Uganda. RESULTS: We tested 1671 samples (123 before ART) with Dynabeads and 1444 samples (91 before ART) with Cytospheres. Mean CD4 was 231 cells/mm (SD, 139) and 239 cells/mm (SD, 140) by Dynabeads and flow cytometry, respectively. Mean CD4 was 186 cells/mm (SD, 101) and 242 cells/mm (SD, 136) by Cytospheres and flow cytometry, respectively. The mean difference in CD4 count by flow cytometry versus Dynabeads were 8.8 cells/mm (SD, 76.0) and versus Cytospheres were 56.8 cells/mm (SD, 85.8). The limits of agreement were -140.9 to 158.4 cells/mm for Dynabeads and -112.2 to 225.8 cells/mm for Cytospheres. Linear regression analysis showed higher correlation between flow cytometry and Dynabeads (r = 0.85, r = 0.73, slope = 0.85, intercept = 28) compared with the correlation between flow cytometry and Cytospheres (r = 0.78, r = 0.60, slope = 0.58, intercept = 45). Area under the receiver operating characteristics curve to predict CD4 <200 cells/mm was 0.928 for Dynabeads and 0.886 for Cytospheres. CONCLUSION: Although Dynabeads and Cytospheres both underestimated CD4 lymphocyte count compared with flow cytometry, in resource-limited settings with low daily throughput, manual bead-based assays may provide a less expensive alternative to flow cytometry.

Original languageEnglish (US)
Pages (from-to)297-303
Number of pages7
JournalJournal of acquired immune deficiency syndromes
Volume48
Issue number3
DOIs
StatePublished - Jul 1 2008

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Keywords

  • Antiretroviral therapy
  • CD4 lymphocyte counts
  • HIV-1 viral load, monitoring and evaluation
  • Resource-limited setting
  • Uganda

ASJC Scopus subject areas

  • Infectious Diseases
  • Pharmacology (medical)

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