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Open Access 01-06-2019 | Acute Myeloid Leukemia | Original Article

A novel flow cytometric method for enhancing acute promyelocytic leukemia screening by multidimensional dot-plots

Authors: Bettina Kárai, Mira Habók, Gyula Reményi, László Rejtő, Anikó Ujfalusi, János Kappelmayer, Zsuzsanna Hevessy

Published in: Annals of Hematology | Issue 6/2019

Abstract

Acute promyelocytic leukemia (APL) is generally characterized by t(15;17)(q24;q21). In some cases, the classic translocation cannot be identified by conventional methods, since the PML-RARA fusion protein results from complex, variant, or cryptic translocation. The diagnostic algorithm of APL starts with screening methods, such as flow cytometry (FC), followed by fluorescence in situ hybridization or polymerase chain reaction to confirm the diagnosis. Our aim was to develop a novel protocol for analyzing APL samples based on multidimensional dot-plots that can provide comprehensive information about several markers at the same time. The protocol included four optimized multidimensional dot-plots, which were tested by retrospective reanalysis of FC results in APL (n = 8) and non-APL (n = 12) acute myeloid leukemia (AML) cases. After predicting the potential position of hypergranular- and microgranular-type aberrant promyelocytes, the percentages of blast populations were examined within the gates in all AML cases. The percentage of blasts in each predefined gate was well above the cut-off value (95%) in APL cases in all tubes. In non-APL AML cases, the percentage of blasts in the same gates never reached the cut-off value in all investigated tubes, and even when it did in a single tube, the pattern was markedly different from that observed in APL cases. In conclusion, multidimensional dot-plots can be used for screening APL even in cryptic APL cases, although reproducibility across several laboratories would require standardization of antibodies and fluorochromes. This easy-to-use and quick method can support the diagnosis of APL and the prompt initiation of the appropriate treatment.

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Title: A novel flow cytometric method for enhancing acute promyelocytic leukemia screening by multidimensional dot-plots
Authors: Bettina Kárai
Mira Habók
Gyula Reményi
László Rejtő
Anikó Ujfalusi
János Kappelmayer
Zsuzsanna Hevessy
Publication date: 01-06-2019
Publisher: Springer Berlin Heidelberg
Keywords: Acute Myeloid Leukemia
Acute Promyelocytic Leukemia
Published in: Annals of Hematology / Issue 6/2019
Print ISSN: 0939-5555
Electronic ISSN: 1432-0584
DOI: https://doi.org/10.1007/s00277-019-03642-w

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