Evaluation of a dual‐color flow cytometry immunophenotyping panel in a multicenter quality assurance program

Eve Levine Schenker, Lance E. Hultin, Kenneth D. Bauer, John Ferbas, Joseph B. Margolick, Janis V. Giorgi

Research output: Contribution to journalArticle

Abstract

A basic immunophenotyping panel that employed dual‐color combinations of fluorescein isothiocyanate (FITC) and phycoerythrin (PE) conjugated monoclonal antibodies (mAb; FITC‐CD45/PE‐CD14, FITC‐IgG1/PE‐IgG2, FITC‐CD3/PE‐CD8, FITC‐CD3/PE‐CD4, FITC‐CD3/PE‐CD16+PE‐CD56, and PE‐CD19) was utilized in a quality assurance program to determine whether the 4 laboratories participating in a multicenter AIDS study obtained similar lymphocyte subset percentage values for T cells, B cells, NK cells, and CD4+ and CD8+ T cells. Over a 1½ year period, 78 shared peripheral blood specimens Were prepared and analyzed in each laboratory. The CD45brightCD14 percentage for each specimen was used to correct that individual's lymphocyte subset values. Interlaboratory coefficients of variation (CV) for the human immunodeficiency virus type I (HIV) seronegative (n = 38) and HIV‐seropositive (n = 40) specimens using this panel were <3% for total T cells; <5% for CD4+ T cells and CD8+ T cells; ≤17% for B and NK cells; and <8% for CD4T/CD8T ratios. The 6‐tube basic immunophenotyping panel has several notable features: (a) for clinical studies, it permits comprehensive evaluation of an individual's major lymphocyte subsets, i.e., T, B, NK, and CD4+ and CD8+ T cells; (b) for interlaboratory proficiency testing programs, it allows the detection of differences among laboratories in measurements of several functionally distinct cell populations; and (c) for within‐sample quality assurance, it provides several quality control checks, including the lym‐phosum, i.e., the sum of an individual's corrected T + B + NK values, a sum that was generally 100 ± 5% on the HIV‐seronegative specimens analyzed in this study. © 1993 Wiley‐Liss, INc.

Original languageEnglish (US)
Pages (from-to)307-317
Number of pages11
JournalCytometry
Volume14
Issue number3
DOIs
StatePublished - 1993

Keywords

  • AIDS
  • Monoclonal antibodies
  • T lymphocyte subsets
  • human immunodeficiency virus type I
  • lymphosum
  • quality control

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

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