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Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia

P. Theunissen, E. Mejstrikova, L. Sedek, AJ. van der Sluijs-Gelling, G. Gaipa, M. Bartels, E. Sobral da Costa, M. Kotrová, M. Novakova, E. Sonneveld, C. Buracchi, P. Bonaccorso, E. Oliveira, JG. Te Marvelde, T. Szczepanski, L. Lhermitte, O....

. 2017 ; 129 (3) : 347-357. [pub] 20161130

Jazyk angličtina Země Spojené státy americké

Typ dokumentu časopisecké články, multicentrická studie

Perzistentní odkaz   https://www.medvik.cz/link/bmc17031253

Grantová podpora
NV15-28525A MZ0 CEP - Centrální evidence projektů

A fully-standardized EuroFlow 8-color antibody panel and laboratory procedure was stepwise designed to measure minimal residual disease (MRD) in B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) patients with a sensitivity of ≤10(-5), comparable to real-time quantitative polymerase chain reaction (RQ-PCR)-based MRD detection via antigen-receptor rearrangements. Leukocyte markers and the corresponding antibodies and fluorochromes were selected based on their contribution in separating BCP-ALL cells from normal/regenerating BCP cells in multidimensional principal component analyses. After 5 multicenter design-test-evaluate-redesign phases with a total of 319 BCP-ALL patients at diagnosis, two 8-color antibody tubes were selected, which allowed separation between normal and malignant BCP cells in 99% of studied patients. These 2 tubes were tested with a new erythrocyte bulk-lysis protocol allowing acquisition of high cell numbers in 377 bone marrow follow-up samples of 178 BCP-ALL patients. Comparison with RQ-PCR-based MRD data showed a clear positive relation between the percentage concordant cases and the number of cells acquired. For those samples with >4 million cells acquired, concordant results were obtained in 93% of samples. Most discordances were clarified upon high-throughput sequencing of antigen-receptor rearrangements and blind multicenter reanalysis of flow cytometric data, resulting in an unprecedented concordance of 98% (97% for samples with MRD < 0.01%). In conclusion, the fully standardized EuroFlow BCP-ALL MRD strategy is applicable in >98% of patients with sensitivities at least similar to RQ-PCR (≤10(-5)), if sufficient cells (>4 × 10(6), preferably more) are evaluated.

Citace poskytuje Crossref.org

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