Background: Flow cytometric analysis of the lymphocyte subsets has become one of the most commonly used techniques in the routine clinical laboratory. It is frequently used in monitoring lymphocyte recovery after hematopoietic stem cell transplantation (HSCT), as well as diagnosis and treatment of acquired immunodeficiency syndrome (AIDS). Reliable biological variation (BV) data is needed for safe clinical application of these tests. In this study, similar preanalytical and analytical protocols to the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) checklist were followed and a stringent statistical approach was applied to define BV of T-lymphocytes.
Methods: During the 10 weeks study period, weekly blood samples were obtained from 30 healthy individuals (20 females, 10 males) and analyzed with Facs Canto (BD Biosciences, San Jose, CA, USA) analyzer using 4-colour BD Multitest CD3/CD8/CD45/CD4 reagents. Data were assessed in terms of normality, tendencies, outliers and variance homogeneity prior to applying coefficient of variance (CV)- analysis of variance (ANOVA) test. Sex-stratified within-individual (CVI) and between-individual (CVG) BV estimates of CD3+, CD3 + CD4+, CD3 + CD8+, and CD3 + CD4 + CD8+ T lymphocytes were calculated.
Results: No difference was found between males and females. Except for the CD3 + CD4 + CD8+ subset, stable BV was found for CD3+, CD3 + CD4+, and CD3 + CD8+ subsets.
Conclussion: Instead of using the conventional reference ranges of CD3+, CD3 + CD4+ and CD3 + CD8+ counts for monitoring HIV positive or post-HSCT patients, RCV should be used. Because individualityis characteristic of lymphocytes subsets RCVs should be used instead of RIs for patient monitoring.
Keywords: Biological variation; Flow cytometry; T lymphocytes.
Published by Elsevier B.V.