title: Evaluation of the FACScount, TRAx CD4 and Dynabeads methods for CD4 lymphocyte determination.
abstract: A study to evaluate the performance of the FACScount, TRAx CD4 and Dynabeads methods for the determination of CD4+ T lymphocyte subset levels was conducted in Tanzania as part of a World Health Organization (WHO) collaborative multicenter field evaluation of alternative methodologies for the enumeration of CD4+ T lymphocytes. The objective was to compare the performance of these alternative methods in a developing country setting, against that of flow cytometry as the reference standard. T lymphocyte subset levels were determined in 91 HIV seronegative and 98 HIV-1 seropositive adults using the three alternative methods. CD4+ and CD8+ T lymphocyte counts were determined by all methods except for TRAx CD4 enzyme linked immunosorbent assay (ELISA) which measures CD4+ T lymphocyte levels only. Linear regression analysis was done to correlate the counts obtained by the alternative methods to those obtained by flow cytometry. The overall correlation coefficients of FACScount and Dynabeads CD4+ and CD8+ T lymphocyte counts with those of flow cytometry were high (r > 0.9). A lower correlation (r = 0.631) was obtained when TRAx CD4+ ELISA counts were compared to those of the reference method. These results show that two of these alternative methodologies are suitable for the determination of CD4+ and CD8+ T lymphocyte counts with the use of African blood samples. Since the methods are simpler and cheaper than flow cytometry, they provide an alternative option for the enumeration of T lymphocyte subsets in laboratories with limited facilities.
authors.affiliation: Department of Microbiology/Immunology, Muhimbili University College of Health Sciences, University of Dar es Salaam, Tanzania.
authors.forename: E F
authors.initials: EF
authors.lastname: Lyamuya
authors.forename: C
authors.initials: C
authors.lastname: Kagoma
authors.forename: E C
authors.initials: EC
authors.lastname: Mbena
authors.forename: W K
authors.initials: WK
authors.lastname: Urassa
authors.forename: K
authors.initials: K
authors.lastname: Pallangyo
authors.forename: F S
authors.initials: FS
authors.lastname: Mhalu
authors.forename: G
authors.initials: G
authors.lastname: Biberfeld
mesh_terms: D000328:Adult; D000349:Africa; D015704:CD4 Antigens / Q000032:analysis; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology*; D002452:Cell Count; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry / Q000379:methods; D006679:HIV Seropositivity / Q000276:immunology; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D014944:World Health Organization
chemical_list: D015704:CD4 Antigens