External quality assurance (EQA) programs are vital to ensure high quality and standardized results in molecular diagnostics. It is important that EQA for quantitative analysis takes into account the variation in methodology. Results cannot be expected to be more accurate than limits of the technology used, and it is essential to recognize factors causing substantial outlier results. The present study aimed to identify parameters of specific importance for JAK2 V617F quantification by quantitative PCR, using different starting materials, assays, and technical platforms. Sixteen samples were issued to participating laboratories in two EQA rounds. In the first round, 19 laboratories from 11 European countries analyzing JAK2 V617F as part of their routine diagnostics returned results from in-house assays. In the second round, 25 laboratories from 17 countries participated. Despite variations in starting material, assay set-up and instrumentation the laboratories were generally well aligned in the EQA program. However, EQA based on a single technology appears to be a valuable tool to achieve standardization of the quantification of JAK2 V617F allelic burden.

Aarhus University Hospital Aarhus Denmark

Belfast City Hospital Belfast UK

CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences Vienna Austria and Department of Internal Medicine 1 Medical University of Vienna Vienna Austria

Center for Biomolecular Pharmaceutical Analyses Faculty of Pharmacy UKiM Skopje Republic of Macedonia

Central Hospital of Southern Pest Budapest Hungary

Centro di Ricerca e Innovazione per le Malattie Mieloproliferative Florence Italy

CHU Côte de Nacre Caen France

CHU de Brest Brest France

CHU Dijon INSERM U866 Dijon France

Clinical Hematology Unit Hospital Pediátrico Centro Hospitalar e Universitário de Coimbra Coimbra Portugal

CRCINA Inserm UMR892 CNRS UMR6299 Centre de Recherche en Cancérologie et Immunologie Nantes Angers Université de Nantes Nantes France

Department of Clinical Chemistry and Transfusion Medicine Institute of Biomedicine the Sahlgrenska Academy University of Gothenburg Gothenburg Sweden

Department of Hematology and Bone Marrow Transplantation Poznan University of Medical Sciences Poznan Poland

Department of Hematology and Department of Pathology Molecular Unit Herlev Hospital University of Copenhagen Herlev Ringvej 75 DK 2730 Herlev Denmark

Department of Hematology Zealand University Hospital Roskilde Denmark

Department of Laboratory Medicine and Pathology Qatar Rehabilitation Institute Doha Qatar

Department of Laboratory Medicine University Hospital of Padova Padova Italy

Department of Pathology Hospital del Mar Barcelona Spain

Department of Pathology Odense University Hospital Odense Denmark

Dubrava University Hospital and Zagreb School of Medicine University of Zagreb Zagreb Croatia

Faculty of Medicine Nicolaus Copernicus University in Torun Bydgoszcz Poland

Haukeland University Hospital Bergen Norway

Hématologie Biologique CHRU de Montpellier Montpellier France

Institute of Hematology and Blood Transfusion Prague Czech Republic

Laboratory of Hematology University Hospital Nantes Nantes France

Molecular Diagnostics Laboratory Hematology Diagnostics Department Jagiellonian University Hospital Krakow Poland

Munich Leukemia Laboratory Munich Germany

National Research Center for Hematology Moscow Russia

Rigshospitalet Copenhagen Denmark

Service de Biologie Cellulaire AP HP Hopital Saint Louis Paris France

St James's Hospital Dublin Ireland

University Hospital Bern and University of Bern Bern Switzerland

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