Original scientific paper

https://doi.org/10.11613/BM.2018.020704

The impact of delayed sample handling and type of anticoagulant on the interpretation of dysplastic signs detected by flow cytometry

Bettina Kárai ; Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Zsófia Miltényi ; Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Lajos Gergely ; Department of Internal Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Marianna Száraz-Széles ; Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
János Kappelmayer ; Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Zsuzsanna Hevessy ; Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary

Abstract

Introduction: A growing body of evidence supports the usefulness of dysplastic signs detected by flow cytometry in the diagnosis of myelodysplastic
syndromes (MDS). Our aim was to assess the impact of pre-analytical variables (delayed sample handling, type of anticoagulant, and different
clones of antibody) in the interpretation of flow cytometric results.
Material and methods: Bone marrow samples were labelled and analysed immediately after aspiration and on two consecutive days. The effect
of anticoagulant type was evaluated in 16 bone marrow samples. Thirty-seven different immunophenotypic variables were recorded after eight-colour
staining. Furthermore, 8 normal peripheral blood samples collected in K3-EDTA and Na-heparin were examined with different clones of CD11b
antibodies and four parameters were recorded with both anticoagulants on two consecutive days.
Results: Fourteen significant differences were detected in the initial immunophenotype of fresh samples collected in K3-EDTA and Na-heparin. Regardless
of the anticoagulant type, eleven parameters remained stable despite delayed sample handling. Due to delayed sample processing, more
alterations were detected in the samples collected in K3-EDTA than in the samples collected in Na-heparin. The type of CD11b clone influenced the
reduction of fluorescence intensity only in samples collected in K3-EDTA, where the alterations were contrary to the changes observed in Na-heparin.
Conclusions: Delayed sample processing causes considerable immunohenotypic alterations, which can lead to false interpretation of the results.
If delayed sample evaluation is unavoidable, markers that remain more stable over time should be considered with more weight in the diagnosis of
MDS.

Keywords

myelodysplastic syndromes; flow cytometry; pre-analytical error

Hrčak ID:

198916

URI

https://hrcak.srce.hr/198916

Publication date:

15.6.2018.

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