Comparison of interlaboratory variation in absolute T-cell counts by single-platform and optimized dual-platform methods.
Abstract
BACKGROUND: Previous studies have reported that the adoption of a single-platform
flow cytometry cell counting method resulted in lower interlaboratory variation in
absolute T cell counts as compared to predicate dual-platform flow cytometry methods
which incorporate independent automated lymphocyte counts (Schnizlein-Bick et al.,
Clin Diagn Lab Immunol 2000;7:336-343; Reimann et al., Clin Diagn Lab Immunol 2000;7:344-351).
In the present study, we asked whether use of a single-platform method could reduce
variation in absolute cell counts across the laboratories in the Multicenter AIDS
Cohort Study (MACS) (n = 4), as suggested by the studies cited. METHODS: Identical
study samples were shipped overnight to the MACS laboratories either by the National
Institute of Allergy and Infectious Diseases, Division of AIDS Immunology Quality
Assessment (NIAID- IQA) proficiency-testing program (n = 14), or by the Los Angeles
site of the MACS (n = 10). For each sample, two tubes of blood were received; one
was used for an automated complete blood count and differential, and the other for
flow cytometry. The latter was performed using both our current dual-platform method
(three-color CD45 gating and automated hematology) and the single-platform method
(with TruCOUNT beads to generate the absolute counts). RESULTS: The median percent
coefficients of variation (%CVs) for the dual-platform and single-platform methods
were 6.6 and 9.9, respectively, for CD4 T cell counts, and 5.9 and 8.5, respectively,
for CD8 T cell counts (n = 24). These differences were not statistically significant.
The differences in absolute T-cell counts between the MACS sites and the median of
all laboratories participating in the NIAID-IQA were smaller for the dual-platform
than for single-platform absolute count method. CONCLUSION: In contrast to previous
reports, we did not observe lower interlaboratory variation across the MACS sites
for single-platform absolute lymphocyte subset counting relative to dual-platform
methods. This result may be at least partly explained by the lower interlaboratory
variation with the optimized dual-platform method in this study relative to the previous
reports.
Type
Journal articleSubject
Cohort StudiesFlow Cytometry
Humans
Laboratories
Lymphocyte Count
Reference Standards
Reproducibility of Results
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https://hdl.handle.net/10161/14742Published Version (Please cite this version)
10.1002/cyto.b.20500Publication Info
Hultin, Lance E; Chow, Marianne; Jamieson, Beth D; O'Gorman, Maurice RG; Menendez,
Frederick A; Borowski, Luann; ... Margolick, Joseph B (2010). Comparison of interlaboratory variation in absolute T-cell counts by single-platform
and optimized dual-platform methods. Cytometry B Clin Cytom, 78(3). pp. 194-200. 10.1002/cyto.b.20500. Retrieved from https://hdl.handle.net/10161/14742.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Thomas Norton Denny
Professor in Medicine
Thomas N. Denny, MSc, M.Phil, is the Chief Operating Officer of the Duke Human Vaccine
Institute (DHVI), Associate Dean for Duke Research and Discovery @RTP, and a Professor
of Medicine in the Department of Medicine at Duke University Medical Center. He is
also an Affiliate Member of the Duke Global Health Institute. Previously, he served
on the Health Sector Advisory Council of the Duke University Fuquay School of Business.
Prior to joining Duke, he was an Associate Professor of Pathology, Labo

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