Development and evaluation of a secondary reference panel for BCR-ABL1 quantification on the International Scale.
Molecular monitoring of chronic myeloid leukemia patients using robust BCR-ABL1 tests standardized to the International Scale (IS) is key to proper disease management, especially when treatment cessation is considered. Most laboratories currently use a time-consuming sample exchange process with reference laboratories for IS calibration. A World Health Organization (WHO) BCR-ABL1 reference panel was developed (MR(1)-MR(4)), but access to the material is limited. In this study, we describe the development of the first cell-based secondary reference panel that is traceable to and faithfully replicates the WHO panel, with an additional MR(4.5) level. The secondary panel was calibrated to IS using digital PCR with ABL1, BCR and GUSB as reference genes and evaluated by 44 laboratories worldwide. Interestingly, we found that >40% of BCR-ABL1 assays showed signs of inadequate optimization such as poor linearity and suboptimal PCR efficiency. Nonetheless, when optimized sample inputs were used, >60% demonstrated satisfactory IS accuracy, precision and/or MR(4.5) sensitivity, and 58% obtained IS conversion factors from the secondary reference concordant with their current values. Correlation analysis indicated no significant alterations in %BCR-ABL1 results caused by different assay configurations. More assays achieved good precision and/or sensitivity than IS accuracy, indicating the need for better IS calibration mechanisms.
Wessex Regional Genetics Laboratory, Salisbury NHS Foundation Trust, Salisbury, UK.|Faculty of Medicine, University of Southampton, Southampton, UK.
N C P
NC
Cross
Wessex Regional Genetics Laboratory, Salisbury NHS Foundation Trust, Salisbury, UK.|Faculty of Medicine, University of Southampton, Southampton, UK.
H E
HE
White
Department of Hematology/Oncology, Universitätsklinikum Jena, Jena, Germany.
T
T
Ernst
Department of Genetics and Molecular Pathology, Centre for Cancer Biology, SA Pathology, Adelaide, SA, Australia.
L
L
Welden
III. Medizinische Klinik, Medizinische Fakultät Mannheim, Universität Heidelberg, Mannheim, Germany.
C
C
Dietz
Department of Clinical and Biological Sciences, San Luigi Hospital, University of Turin, Orbassano, Italy.
G
G
Saglio
Bergonie Institute Cancer Center Bordeaux, INSERM U1218, University of Bordeaux, Bordeaux, France.
F-X
FX
Mahon
Novartis Pharmaceuticals Corporation, East Hanover, NJ, USA.
C C
CC
Wong
Novartis Pharmaceuticals Corporation, East Hanover, NJ, USA.
D
D
Zheng
Novartis Pharmaceuticals Corporation, East Hanover, NJ, USA.
S
S
Wong
Novartis Pharmaceuticals Corporation, East Hanover, NJ, USA.
S-S
SS
Wang
West Midlands Regional Genetics Laboratory, Birmingham, UK.
S
S
Akiki
Department of Hematology, University of Bari, Bari, Italy.
F
F
Albano
Laboratory of Molecular Diagnostics, Hungarian National Blood Transfusion Service, Budapest, Hungary.|Department of Pathophysiology, Semmelweis University, Budapest, Hungary.
H
H
Andrikovics
King's College Hospital London, London, UK.
J
J
Anwar
National Specialized Hospital for Active Treatment of Hematological Diseases, Sofia, Bulgaria.
G
G
Balatzenko
Laboratorio de Biologia Tumoral, Disciplina de Hematologia do HC-FMUSP, São Paulo, Brazil.
I
I
Bendit
PathWest Laboratory Medicine WA, Department of Haematology, Fiona Stanley Hospital, Perth, WA, Australia.
J
J
Beveridge
Department of Laboratory Medicine, University Hospitals Leuven, Leuven, Belgium.|Department of Oncology, KUL, Leuven, Belgium.
N
N
Boeckx
Department of Genetics, Portuguese Oncology Institute, Porto, Portugal.
N
N
Cerveira
Department of Hematology and Oncology, Quest Diagnostics Nichols Institute, San Juan Capistrano, CA, USA.
S-M
SM
Cheng
Hematopathology Unit, Hospital Clinic, IDIBAPS, Barcelona, Spain.
D
D
Colomer
Division of Molecular Microbiology, Children's Cancer Research Institute, Vienna, Austria.
S
S
Czurda
Department of Clinical and Biological Sciences, San Luigi Hospital, University of Turin, Orbassano, Italy.
F
F
Daraio
Laboratoire Hematologie, Centre Hospitalier Universitaire de Bordeaux, Universite Bordeaux, Bordeaux, France.
S
S
Dulucq
Laboratory of Molecular Pathology, Oslo University Hospital, Oslo, Norway.
L
L
Eggen
Clinique de Genetique Oncologique-Service de genetique, Hopital Erasme, Brussels, Belgium.
H
H
El Housni
Imperial Molecular Pathology, Hammersmith Hospital, London, UK.
G
G
Gerrard
Department of Hematology and Bone Marrow Transplantation, Poznan University of Medical Sciences, Poznan, Poland.
M
M
Gniot
Department of Clinical Medicine and Surgery, University 'Federico II' of Naples, Naples, Italy.|CEINGE - Biotecnologie Avanzate, Naples, Italy.
B
B
Izzo
Genoptix, Inc., Carlsbad, CA, USA.
D
D
Jacquin
Department of Hematology and Molecular Diagnostics, VU University Medical Center, Amsterdam, The Netherlands.
J J W M
JJ
Janssen
MLL Munich Leukemia Laboratory, Munich, Germany.
S
S
Jeromin
Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, Masaryk University and University Hospital Brno, Brno, Czech Republic.
T
T
Jurcek
Seoul St Mary's Hospital, Leukemia Research Institute, The Catholic University of Korea, Seoul, Korea.
D-W
DW
Kim
Department of Molecular Genetics, Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
K
K
Machova-Polakova
Department of Hematology, Hospital Universitario 12 de Octubre, Universidad Complutense, CNIO, Madrid, Spain.
J
J
Martinez-Lopez
Division of Cancer Medicine, Department of Pathology, Peter MacCallum Cancer Centre, East Melbourne, VIC, Australia.
M
M
McBean
Pathology Department, University Clinical Center Tuzla, Tuzla, Bosnia and Herzegovina.
S
S
Mesanovic
Department of Laboratory Medicine, Division of Medical and Chemical Laboratory Diagnostics, Medical University of Vienna, Vienna, Austria.
G
G
Mitterbauer-Hohendanner
Genoptix, Inc., Carlsbad, CA, USA.
H
H
Mobtaker
Departement de Biopathologie, Institut Paoli-Calmettes, Marseille, France.
M-J
MJ
Mozziconacci
Specialized Haematology Laboratory, Department of Haematology, University Medical Centre Ljubljana, Ljubljana, Slovenia.
T
T
Pajič
Department of Surgical Pathology, Zealand University Hospital, Roskilde, Denmark.
N
N
Pallisgaard
Hematology Unit, First Department of Internal Medicine, Laiko Hospital, University of Athens, Athens, Greece.
P
P
Panagiotidis
Department of Pathology and Knight Cancer Institute, Oregon Health and Science University, Portland, OR, USA.
R D
RD
Press
Peking University People's Hospital, Peking University Institute of Hematology, Beijing, China.
Y-Z
YZ
Qin
Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
J
J
Radich
Chair and Department of Hematology, Jagiellonian University, Kraków, Poland.
T
T
Sacha
Laboratory of Molecular Biology, Hematology Department and HCT Unit, G. Papanicolaou Hospital, Thessaloniki, Greece.
T
T
Touloumenidou
Bristol Genetics Laboratory, Bristol, UK.
P
P
Waits
HMDS, Leeds Teaching Hospitals, Leeds, UK.
E
E
Wilkinson
Faculty of Pharmacy and Biochemistry and University Hospital Center Zagreb, University of Zagreb, Zagreb, Croatia.
R
R
Zadro
III. Medizinische Klinik, Medizinische Fakultät Mannheim, Universität Heidelberg, Mannheim, Germany.
M C
MC
Müller
Department of Hematology/Oncology, Universitätsklinikum Jena, Jena, Germany.
A
A
Hochhaus
Department of Genetics and Molecular Pathology, Centre for Cancer Biology, SA Pathology, Adelaide, SA, Australia.|School of Pharmacy and Medical Science, University of South Australia, Adelaide, SA, Australia.|School of Medicine, University of Adelaide, SA, Adelaide, Australia.|School of Molecular and Biomedical Science, University of Adelaide, Adelaide, SA, Australia.
S
S
Branford
D002138:Calibration; D016044:Fusion Proteins, bcr-abl / Q000032:analysis* / Q000592:standards; D016313:Genes, abl; D006801:Humans; D016133:Polymerase Chain Reaction; D051562:Proto-Oncogene Proteins c-bcr / Q000235:genetics; D012015:Reference Standards; D014944:World Health Organization
D016044:Fusion Proteins, bcr-abl; C496613:BCR protein, human; D051562:Proto-Oncogene Proteins c-bcr