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Flow cell sorting followed by PCR‐based clonality testing may assist in questionable diagnosis and monitoring of acute lymphoblastic leukemia.

Authors :
Semchenkova, Alexandra
Zhogov, Vladimir
Zakharova, Elena
Mikhailova, Ekaterina
Illarionova, Olga
Larin, Sergey
Novichkova, Galina
Karachunskiy, Alexander
Maschan, Michael
Popov, Alexander
Source :
International Journal of Laboratory Hematology. Aug2023, Vol. 45 Issue 4, p506-515. 10p.
Publication Year :
2023

Abstract

Introduction: Multicolor flow cytometry (MFC) has highly reliable and flexible algorithms for diagnosis and monitoring of acute lymphoblastic leukemia (ALL). However, MFC analysis can be affected by poor sample quality or novel therapeutic options (e.g., targeted therapies and immunotherapy). Therefore, an additional confirmation of MFC data may be needed. We propose a simple approach for validation of MFC findings in ALL by sorting questionable cells and analyzing immunoglobulin/T‐cell receptor (IG/TR) gene rearrangements via EuroClonality‐based multiplex PCR. Patients and Methods: We obtained questionable MFC results for 38 biological samples from 37 patients. In total, 42 cell populations were isolated by flow cell sorting for downstream multiplex PCR. Most of the patients (n = 29) had B‐cell precursor ALL and were investigated for measurable residual disease (MRD); 79% of them received CD19‐directed therapy (blinatumomab or CAR‐T). Results: We established the clonal nature of 40 cell populations (95.2%). By using this technique, we confirmed very low MRD levels (<0.01% MFC‐MRD). We also applied it to several ambiguous findings for diagnostic samples, including those with mixed‐phenotype acute leukemia, and the results obtained impacted the final diagnosis. Conclusion: We have demonstrated possibilities of a combined approach (cell sorting and PCR‐based clonality assessment) to validate MFC findings in ALL. The technique is easy to implement in diagnostic and monitoring workflows, as it does not require isolation of a large number of cells and knowledge of individual clonal rearrangements. We believe it provides important information for further treatment. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
17515521
Volume :
45
Issue :
4
Database :
Academic Search Index
Journal :
International Journal of Laboratory Hematology
Publication Type :
Academic Journal
Accession number :
164960624
Full Text :
https://doi.org/10.1111/ijlh.14053