Assessment of clonality, marker identification and measurement of minimal residual disease (MRD) of immunoglobulin (IG) and T cell receptor (TR) gene rearrangements in lymphoid neoplasms using next-generation sequencing (NGS) is currently under intensive development for use in clinical diagnostics. So far, however, there is a lack of suitable quality control (QC) options with regard to standardisation and quality metrics to ensure robust clinical application of such approaches. The EuroClonality-NGS Working Group has therefore established two types of QCs to accompany the NGS-based IG/TR assays. First, a central polytarget QC (cPT-QC) is used to monitor the primer performance of each of the EuroClonality multiplex NGS assays; second, a standardised human cell line-based DNA control is spiked into each patient DNA sample to work as a central in-tube QC and calibrator for MRD quantification (cIT-QC). Having integrated those two reference standards in the ARResT/Interrogate bioinformatic platform, EuroClonality-NGS provides a complete protocol for standardised IG/TR gene rearrangement analysis by NGS with high reproducibility, accuracy and precision for valid marker identification and quantification in diagnostics of lymphoid malignancies.

Bristol Genetics Laboratory Southmead Hospital Bristol UK

Central European Institute of Technology Masaryk University Brno Czech Republic

Centre for Cancer Research and Cell Biology Queen's University Belfast Belfast UK

Centro Ricerca Tettamanti University of Milano Bicocca Monza Italy

CLIP Childhood Leukaemia Investigation Prague Department of Paediatric Haematology and Oncology 2nd Faculty of Medicine Charles University University Hospital Motol Prague Czech Republic

Department of Hematology APHP Necker Enfants Malades and Paris Descartes University Paris France

Department of Hematology Hopital Pitié Salpêtrière Paris France

Department of Hematology University Hospital Schleswig Holstein Kiel Germany

Department of Immunohematology and Blood Transfusion Leiden University Medical Center Leiden The Netherlands

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

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

Department of Paediatric Haematology Great Ormond Street Hospital London UK

Department of Pathology Radboud University Medical Center Nijmegen The Netherlands

Department of Pediatric Haematology Bristol Royal Hospital for Children Bristol UK

IBMCC CSIC Hospital Universitario de Salamanca IBSAL Salamanca Spain

Insititute of Pathology Charité Universitätsmedizin Berlin Berlin Germany

Institute of Applied Biosciences Centre for Research and Technology Hellas Thessaloniki Greece

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Insights into IGH clonal evolution in BCP-ALL: frequency, mechanisms, associations, and diagnostic implications

NGS better discriminates true MRD positivity for the risk stratification of childhood ALL treated on an MRD-based protocol

IGH Rearrangement Evolution in Adult KMT2A-rearranged B-cell Precursor ALL: Implications for Cell-of-origin and MRD Monitoring

Quality Control for IG /TR Marker Identification and MRD Analysis

Potential and pitfalls of whole transcriptome-based immunogenetic marker identification in acute lymphoblastic leukemia; a EuroMRD and EuroClonality-NGS Working Group study

Standardized next-generation sequencing of immunoglobulin and T-cell receptor gene recombinations for MRD marker identification in acute lymphoblastic leukaemia; a EuroClonality-NGS validation study

Next-generation sequencing of immunoglobulin gene rearrangements for clonality assessment: a technical feasibility study by EuroClonality-NGS