Presence of minimal residual disease (MRD), detected by flow cytometry, is an important prognostic biomarker in the management of B-cell precursor acute lymphoblastic leukemia (BCP-ALL). However, data-analysis remains mainly expert-dependent. In this study, we designed and validated an Automated Gating & Identification (AGI) tool for MRD analysis in BCP-ALL patients using the two tubes of the EuroFlow 8-color MRD panel. The accuracy, repeatability, and reproducibility of the AGI tool was validated in a multicenter study using bone marrow follow-up samples from 174 BCP-ALL patients, stained with the EuroFlow BCP-ALL MRD panel. In these patients, MRD was assessed both by manual analysis and by AGI tool supported analysis. Comparison of MRD levels obtained between both approaches showed a concordance rate of 83%, with comparable concordances between MRD tubes (tube 1, 2 or both), treatment received (chemotherapy versus targeted therapy) and flow cytometers (FACSCanto versus FACSLyric). After review of discordant cases by additional experts, the concordance increased to 97%. Furthermore, the AGI tool showed excellent intra-expert concordance (100%) and good inter-expert concordance (90%). In addition to MRD levels, also percentages of normal cell populations showed excellent concordance between manual and AGI tool analysis. We conclude that the AGI tool may facilitate MRD analysis using the EuroFlow BCP-ALL MRD protocol and will contribute to a more standardized and objective MRD assessment. However, appropriate training is required for the correct analysis of MRD data.

Biomedical Research Networking Centre Consortium of Oncology Instituto de Salud Carlos 3 Madrid Spain

Centro Tettamanti Fondazione IRCCS San Gerardo dei Tintori Monza Italy

CLIP Department of Pediatric Hematology and Oncology 2nd Faculty of Medicine Charles University and University Hospital Motol Prague Czech Republic

Cytognos SL Salamanca Spain

Department of Diagnostic Sciences Ghent University Ghent Belgium

Department of Hematology University of Schleswig Holstein Kiel Germany

Department of Immunology Leiden University Medical Center The Netherlands

Department of Laboratory Medicine Ghent University Hospital Ghent Belgium

Department of Medicine University of Salamanca Salamanca Spain

Department of Microbiology and Immunology Medical University of Silesia in Katowice Zabrze Poland

Department of Pediatric Hematology and Oncology Zabrze Medical University of Silesia in Katowice Katowice Poland

Institute for Laboratory Medicine Aarau Switzerland

Institute of Biomedical Research of Salamanca Salamanca Spain

Laboratory for Medical Immunology Department of Immunology Erasmus MC University Medical Center Rotterdam Rotterdam The Netherlands

Pediatrics Institute IPPMG Faculty of Medicine Federal University of Rio de Janeiro Rio de Janeiro Brazil

Princess Máxima Center for Pediatric Oncology Utrecht The Netherlands

Translational and Clinical Research program Cancer Research Centre Cytometry Service NUCLEUS Salamanca Spain

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