Serial quantification of BCR-ABL1 mRNA is an important therapeutic indicator in chronic myeloid leukaemia, but there is a substantial variation in results reported by different laboratories. To improve comparability, an internationally accepted plasmid certified reference material (CRM) was developed according to ISO Guide 34:2009. Fragments of BCR-ABL1 (e14a2 mRNA fusion), BCR and GUSB transcripts were amplified and cloned into pUC18 to yield plasmid pIRMM0099. Six different linearised plasmid solutions were produced with the following copy number concentrations, assigned by digital PCR, and expanded uncertainties: 1.08±0.13 × 10(6), 1.08±0.11 × 10(5), 1.03±0.10 × 10(4), 1.02±0.09 × 10(3), 1.04±0.10 × 10(2) and 10.0±1.5 copies/μl. The certification of the material for the number of specific DNA fragments per plasmid, copy number concentration of the plasmid solutions and the assessment of inter-unit heterogeneity and stability were performed according to ISO Guide 35:2006. Two suitability studies performed by 63 BCR-ABL1 testing laboratories demonstrated that this set of 6 plasmid CRMs can help to standardise a number of measured transcripts of e14a2 BCR-ABL1 and three control genes (ABL1, BCR and GUSB). The set of six plasmid CRMs is distributed worldwide by the Institute for Reference Materials and Measurements (Belgium) and its authorised distributors (https://ec.europa.eu/jrc/en/reference-materials/catalogue/; CRM code ERM-AD623a-f).

] Department of Laboratory Medicine UZ Leuven Belgium [2] Department of Oncology KU Leuven Belgium

] National Genetics Reference Laboratory Salisbury District Hospital Salisbury UK [2] Faculty of Medicine University of Southampton Southampton UK

3 Medizinische Klinik Medizinische Fakultät Mannheim der Universität Heidelberg Mannheim Germany

Abteilung für Hämatologie und internistische Onkologie Universität Leipzig Leipzig Germany

Bristol Genetics Laboratory Southmead Hospital Bristol UK

Cancer Molecular Diagnostics St James's Hospital Dublin Ireland

Cancer Research Institute The Catholic University of Korea Seoul South Korea

Children's Cancer Research Institute LabDia Labordiagnostik and Medical University Vienna Austria

Combined Laboratories Derriford Hospital Plymouth UK

Department of Clinical and Biological Science University of Turin Turin Italy

Department of Clinical Genetics University and Regional Laboratories Lund Sweden

Department of Clinical Medicine and Surgery University 'Federico II' of Naples Naples Italy

Department of Experimental Diagnostic and Specialty Medicine University of Bologna Bologna Italy

Department of Genetics and Molecular Pathology SA Pathology Adelaide SA Australia

Department of Haematology Aarhus University Hospital Aarhus Denmark

Department of Haematology Royal Liverpool University Hospital Liverpool UK

Department of Hematology Oncology Jena University Hospital Jena Germany

Department of Immunology Erasmus MC Rotterdam The Netherlands

Department of Immunology Genetics and Pathology Uppsala University Uppsala Sweden

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

Department of Internal Medicine Hematology Oncology Goethe University Frankfurt Germany

Department of Laboratory Diagnostics Clinical Hospital Center Zagreb University School of Medicine Zagreb Croatia

Department of Laboratory Medicine Medical University of Vienna Vienna Austria

Department of Medical Genetics NHS Grampian Aberdeen UK

Department of Molecular Haematology Yorkhill NHS Trust Glasgow UK

Department of Molecular Medicine and Surgery Clinical Genetics Karolinska Institutet Stockholm Sweden

Division of Pathology Rikshospital Oslo University Hospital Oslo Norway

European Commission Joint Research Centre Institute for Reference Materials and Measurements Geel Belgium

Genetics Department Institute of Experimental Medicine Istanbul University Istanbul Turkey

Haematology Department Belfast City Hospital Belfast UK

Haematology Laboratory and EA3518 University Hospital Saint Louis AP HP University Paris Diderot Paris France

Haematology Research Laboratory Biomedical Research Foundation Academy of Athens Athens Greece

Hematology Department and HCT Unit G Papanicolaou Hospital Thessaloniki Greece

Hematology Department Fundeni Clinical Institute University of Medicine and Pharmacy 'Carol Davila' Bucharest Romania

Hematology Department Jagiellonian University Krakow Poland

Hematology Oncology and Transfusion Medicine Center Vilnius University Hospital Santariskiu Clinics Vilnius Lithuania

Hematopathology Unit Hospital Clinic IDIBAPS Barcelona Spain

HMDS Leeds Institute of Oncology St James's University Hospital Leeds UK

Hungarian National Blood Transfusion Service Budapest Hungary

Imperial Molecular Pathology Centre for Haematology Imperial College London London UK

Institute of Hematology and Blood Transfusion Prague Czech Republic

Lab Hematologia Hospital de la Santa Creu i Sant Pau Barcelona Spain

Laboratoire d'hématologie CHU Lille Lille France

Laboratoire Hematologie CHU Bordeaux Hematopoiese Leucemique et Cibles Therapeutiques INSERM U1035 Universite Bordeaux Bordeaux France

Laboratory for Molecular Haemato Oncology Kings College Hospital London UK

Laboratory of Hematology Sheba Medical Center Tel Hashomer Israel

Laboratory of Molecular Biology and UMR5239 Centre Hospitalier Lyon Sud Hospices Civils de Lyon Pierre Bénite France

Medical Genetics Department Erasme Hospital Brussels Belgium

Molecular Diagnostics Laboratory Department of Hematology University Hospital Bern Bern Switzerland

Molecular Diagnostics The Royal Marsden NHS Foundation Trust Sutton UK

Molecular Haematology Bristol Royal Infirmary Bristol UK

Molecular Haematology Laboratory PathWest Laboratory Medicine Royal Perth Hospital Perth WA Australia

Molecular Malignancy Laboratory and Haemato Oncology Diagnostic Service Cambridge University Hospitals NHS Foundation Trust Cambridge UK

Molecular Oncology Diagnostics Unit Guy's Hospital London UK

Molecular Pathology University Hospitals Southampton NHS Foundation Trust Southampton UK

National Genetics Reference Laboratory Salisbury District Hospital Salisbury UK

Regional Genetics Laboratory Birmingham Women's NHS Foundation Trust Birmingham UK

Sheffield Diagnostic Genetics Service Sheffield Children's NHS Foundation Trust Sheffield UK

Specialized Haematology Laboratory Division of Internal Medicine Department of Haematology University Medical Centre Ljubljana Slovenia

Turku University Hospital TYKSLAB Laboratory of Molecular Genetics Turku Finland

United Laboratories of Tartu University Hospitals Tartu Estonia

VU Medical Centre Department of Haematology Amsterdam The Netherlands

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European LeukemiaNet laboratory recommendations for the diagnosis and management of chronic myeloid leukemia

Standardization of molecular monitoring of CML: results and recommendations from the European treatment and outcome study

Impact of BCR::ABL1 transcript type on RT-qPCR amplification performance and molecular response to therapy

International external quality assurance of JAK2 V617F quantification

Proteomic analysis of imatinib-resistant CML-T1 cells reveals calcium homeostasis as a potential therapeutic target

Laboratory recommendations for scoring deep molecular responses following treatment for chronic myeloid leukemia