We present the results of a pilot study concerning the interlaboratory variability of CD34+ enumeration. Three surveys, each including a set of samples, were sent to participating Czech flow cytometry laboratories. The efficacy of this exercise was determined by the reduction in interlaboratory variation and the influence of method used on assay outcome. The variability in results of CD34+ enumeration declined with time. The mean coefficient of variation (CV) of measurement among laboratories dropped, from 58% in first survey to 32% in last survey. All tested variables (gating strategy, platform methodology, sample preparation) affected the variability of the assay. Sample preparation method was associated with a significant bias of absolute CD34+ cell counts. Initially, the outcome of the measurement was also affected by the participating laboratory (identified by a unique laboratory number; ULN). However, laboratories with poorer performance modified their protocols during the study, and the ULN ceased to influence the variability. This study was successful in reducing the interinstitutional variability of CD34+ enumeration. It was shown that the implementation of a standardized protocol does not guarantee accurate measurement. Our research design represents a useful tool, which allows verification of the proper use of a standardized method, the training of operators and feedback in response to the survey results.
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