A single-tube, 20-color panel effectively detects aberrant cells in acute myeloid leukemia.

Abstract

Abstract Objective: Multiparameter flow cytometry is widely and routinely used in acute myeloid leukemia (AML) diagnosis and the residual disease detection and monitoring. We have developed a single-tube, 20-color panel for AML analysis that uses the Cytek® spectral flow cytometer with increased reagent and sample efficiency. Methods: Blood and bone marrow samples were stained with the 20-color panel, acquired, and analyzed using SpectroFlo® software. The resolution of each marker was compared between single stain versus the 20-color full stain on the same samples. The analytical precision for cell populations of interest and antibody cocktail stability were assessed. The limit of detection (LOD) and lower limit of quantification (LLOQ) of the 20-color assay were determined using limiting dilution method. Results: The resolution of each marker in the fully stained panel was comparable to that in the single-color stained samples. Normal and aberrant myeloid cell populations were clearly identified and the coefficient of variation (CV) of cell percentage for defined populations in replicate runs were all less than 25%. The antibody cocktail was stable in the fridge for at least 14 days. Our data showed that the single-tube, 20-color assay easily achieved 0.01% on residual aberrant cells in AML, higher than the minimum required detection sensitivity of 0.1%. Conclusions: Cytek’s single-tube, 20-color panel demonstrated an effective, high sensitivity flow cytometry approach that can be used for AML testing, including identifying and characterizing normal and aberrant cells, immunophenotypic classification and minimal residual disease evaluation in translation research.