Evaluation of a dual-color flow cytometry immunophenotyping panel in a multicenter quality assurance program.
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 1 1/2 year period, 78 shared peripheral blood specimens were prepared and analyzed in each laboratory. The CD45bright CD14- 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; < or = 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 lymphosum, 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.
Schenker
E L
EL
Department of Medicine, UCLA School of Medicine 90024.
Hultin
L E
LE
Bauer
K D
KD
Ferbas
J
J
Margolick
J B
JB
Giorgi
J V
JV
D015492:AIDS Serodiagnosis / Q000379:methods*; D000163:Acquired Immunodeficiency Syndrome / Q000097:blood; D000911:Antibodies, Monoclonal / Q000276:immunology; D016175:B-Lymphocyte Subsets / Q000276:immunology*; D016516:CD4-CD8 Ratio; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D010799:Phycoerythrin; D011786:Quality Control; D016176:T-Lymphocyte Subsets / Q000276:immunology*
D000911:Antibodies, Monoclonal; D010799:Phycoerythrin; D016650:Fluorescein-5-isothiocyanate