Reduction of variation in T‐cell subset enumeration among 55 laboratories using single‐platform, three or four‐color flow cytometry based on CD45 and SSC‐based gating of lymphocytes

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Abstract
BACKGROUND Enumeration of CD4+ and CD8+ T‐cell subsets provides relevant information for diagnosis and monitoring of patients with cellular immunodeficiencies. As a result, an external quality assurance scheme was implemented in Belgium, The Netherlands, and Luxemburg in 1995. A workshop was held to train the participants in state‐of‐the art technology for assessment of absolute T‐cell subset counts (i.e., a three or four‐color, single‐platform assay with lymphocyte gating based on CD45 and sideward light scatter) with the aim to achieve between‐site coefficients of variation (CVs) + T cells), ∼9% (CD4+ T cells), and ∼10% (CD8+ T cells) were achieved. Within‐site CVs were + T cells) and ∼70% (CD4+ and CD8+ subsets) of the participants. Local techniques yielded between‐site CVs of 13%–17% for CD3+, CD4+, and CD8+ T cells. Conclusions: The state‐of‐the‐art technology for T‐cell subset enumeration was implemented successfully among 55 Belgian‐Dutch laboratories and resulted in significant reductions of between‐site variation of absolute CD3+, CD4+, and CD8+ T‐cell counts. Cytometry (Clin. Cytometry) 50:92–101, 2002.

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