title: Flow cytometric immunobead assay for fast and easy detection of PML-RARA fusion proteins for the diagnosis of acute promyelocytic leukemia.
abstract: The PML-RARA fusion protein is found in approximately 97% of patients with acute promyelocytic leukemia (APL). APL can be associated with life-threatening bleeding complications when undiagnosed and not treated expeditiously. The PML-RARA fusion protein arrests maturation of myeloid cells at the promyelocytic stage, leading to the accumulation of neoplastic promyelocytes. Complete remission can be obtained by treatment with all-trans-retinoic acid (ATRA) in combination with chemotherapy. Diagnosis of APL is based on the detection of t(15;17) by karyotyping, fluorescence in situ hybridization or PCR. These techniques are laborious and demand specialized laboratories. We developed a fast (performed within 4-5 h) and sensitive (detection of at least 10% malignant cells in normal background) flow cytometric immunobead assay for the detection of PML-RARA fusion proteins in cell lysates using a bead-bound anti-RARA capture antibody and a phycoerythrin-conjugated anti-PML detection antibody. Testing of 163 newly diagnosed patients (including 46 APL cases) with the PML-RARA immunobead assay showed full concordance with the PML-RARA PCR results. As the applied antibodies recognize outer domains of the fusion protein, the assay appeared to work independently of the PML gene break point region. Importantly, the assay can be used in parallel with routine immunophenotyping for fast and easy diagnosis of APL.
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
authors.forename: E H A
authors.initials: EH
authors.lastname: Dekking
authors.forename: V H J
authors.initials: VH
authors.lastname: van der Velden
authors.forename: R
authors.initials: R
authors.lastname: Varro
authors.forename: H
authors.initials: H
authors.lastname: Wai
authors.forename: S
authors.initials: S
authors.lastname: Böttcher
authors.forename: M
authors.initials: M
authors.lastname: Kneba
authors.forename: E
authors.initials: E
authors.lastname: Sonneveld
authors.forename: A
authors.initials: A
authors.lastname: Koning
authors.forename: N
authors.initials: N
authors.lastname: Boeckx
authors.forename: N
authors.initials: N
authors.lastname: Van Poecke
authors.forename: P
authors.initials: P
authors.lastname: Lucio
authors.forename: A
authors.initials: A
authors.lastname: Mendonça
authors.forename: L
authors.initials: L
authors.lastname: Sedek
authors.forename: T
authors.initials: T
authors.lastname: Szczepański
authors.forename: T
authors.initials: T
authors.lastname: Kalina
authors.forename: V
authors.initials: V
authors.lastname: Kanderová
authors.forename: P
authors.initials: P
authors.lastname: Hoogeveen
authors.forename: J
authors.initials: J
authors.lastname: Flores-Montero
authors.forename: M C
authors.initials: MC
authors.lastname: Chillón
authors.forename: A
authors.initials: A
authors.lastname: Orfao
authors.forename: J
authors.initials: J
authors.lastname: Almeida
authors.forename: P
authors.initials: P
authors.lastname: Evans
authors.forename: M
authors.initials: M
authors.lastname: Cullen
authors.forename: A L
authors.initials: AL
authors.lastname: Noordijk
authors.forename: P M
authors.initials: PM
authors.lastname: Vermeulen
authors.forename: M T
authors.initials: MT
authors.lastname: de Man
authors.forename: E P
authors.initials: EP
authors.lastname: Dixon
authors.forename: W M
authors.initials: WM
authors.lastname: Comans-Bitter
authors.forename: J J M
authors.initials: JJ
authors.lastname: van Dongen
mesh_terms: D000328:Adult; D016022:Case-Control Studies; D002648:Child; D002884:Chromosomes, Human, Pair 15 / Q000235:genetics; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007118:Immunoassay; D015473:Leukemia, Promyelocytic, Acute / Q000175:diagnosis* / Q000276:immunology / Q000378:metabolism*; D008297:Male; D015514:Oncogene Proteins, Fusion / Q000276:immunology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
chemical_list: D015514:Oncogene Proteins, Fusion; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein