Assessing agreement between CD34 enumeration by flow cytometry and volumetric analysis

Prior to replacement of an established method for CD34 enumeration by an alternative approach, evaluation of the agreement between the methods is essential. In this study, the comparison of two assays was evaluated according to the recommendation of Bland and Altman describing the agreement between two methods where the true value is not known. CD34 enumeration was performed on blood or leukapheresis product from 105 patients by flow cytometry (dual platform assay) and volumetric analysis (single platform assay). Both the flow cytometric and the volumetric analysis showed poor reproducibility for measures lower than approximately 9 CD34(+) cells/mm(3). For values higher than 29 CD34(+) cells/mm(3), evaluation of the agreement demonstrated a difference between the single and dual platform assay, where CD34 enumeration by the volumetric analysis demonstrated values 73-80% of the flow cytometric value. The difference between the two assays could be due to several technical pitfalls which are discussed.