The diagnostic criteria for CLL rely on morphology and immunophenotype. Current approaches have limitations affecting reproducibility and there is no consensus on the role of new markers. The aim of this project was to identify reproducible criteria and consensus on markers recommended for the diagnosis of CLL. ERIC/ESCCA members classified 14 of 35 potential markers as "required" or "recommended" for CLL diagnosis, consensus being defined as >75% and >50% agreement, respectively. An approach to validate "required" markers using normal peripheral blood was developed. Responses were received from 150 participants with a diagnostic workload >20 CLL cases per week in 23/150 (15%), 5-20 in 82/150 (55%), and <5 cases per week in 45/150 (30%). The consensus for "required" diagnostic markers included: CD19, CD5, CD20, CD23, Kappa, and Lambda. "Recommended" markers potentially useful for differential diagnosis were: CD43, CD79b, CD81, CD200, CD10, and ROR1. Reproducible criteria for component reagents were assessed retrospectively in 14,643 cases from 13 different centers and showed >97% concordance with current approaches. A pilot study to validate staining quality was completed in 11 centers. Markers considered as "required" for the diagnosis of CLL by the participants in this study (CD19, CD5, CD20, CD23, Kappa, and Lambda) are consistent with current diagnostic criteria and practice. Importantly, a reproducible approach to validate and apply these markers in individual laboratories has been identified. Finally, a consensus "recommended" panel of markers to refine diagnosis in borderline cases (CD43, CD79b, CD81, CD200, CD10, and ROR1) has been defined and will be prospectively evaluated. © 2017 International Clinical Cytometry Society.

CEITEC Masaryk University Brno Czech Republic

Department of Clinical Medicine Aalborg University Aalborg Denmark

Department of Clinical Pathology Flow Cytometry Laboratory C H L O Hospital S Francisco Xavier Lisbon Portugal

Department of Haematology Clinical Cancer Research Centre Aalborg University Hospital Aalborg Denmark

Department of Hematology Hospital Clinic University of Barcelona Barcelona Spain

Department of Immunology Genetics and Pathology Science for Life Laboratory Uppsala University Uppsala Sweden

Department of Internal Medicine Hematology and Oncology University Hospital Brno Brno Czech Republic

Dubrava University Hospital Zagreb Croatia

Evangelismos Hospital Athens Greece

Hematopathology Unit Hospital Clínic Barcelona Spain

Hospital de Sant Pau Barcelona Spain

Hospital Universitario 12 de Octubre Madrid Spain

Laverty Pathology Sydney Australia

MLL Munich Leukemia Laboratory Munich Germany

Ospedale San Raffaele Milano Italy

Peter MacCallum Cancer Centre Melbourne Australia

Rigshospitalet Copenhagen University Hospital Copenhagen Denmark

Royal Bournemouth Hospital Bournemouth UK

St Anna University Hospital Ferrara Italy

St James's Institute of Oncology Leeds UK

Università Vita Salute San Raffaele and IRCCS Istituto Scientifico San Raffaele Milano Italy

Universitätsklinikum Köln Köln Germany

University Hospital Prague Czech Republic

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The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours: Lymphoid Neoplasms

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Expert-independent classification of mature B-cell neoplasms using standardized flow cytometry: a multicentric study