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Título

Flow cytometric immunobead assay for fast and easy detection of PML-RARA fusion proteins for the diagnosis of acute promyelocytic leukemia

AutorDekking, E.; Flores-Montero, Juan; Chillón, M. del Carmen; Orfao, Alberto CSIC ORCID ; Almeida, Julia CSIC ORCID CVN
Palabras claveAPL
PML–RARA protein
t(15 17)
Immunobead
Flow cytometry
Fecha de publicación2012
EditorNature Publishing Group
CitaciónLeukemia 26(9): 1976-1985 (2012)
ResumenThe 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.
DescripciónThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivative Works 3.0 Unported License.-- EuroFlow Consortium: et al.
Versión del editorhttp://dx.doi.org/10.1038/leu.2012.125
URIhttp://hdl.handle.net/10261/134149
DOI10.1038/leu.2012.125
Identificadoresdoi: 10.1038/leu.2012.125
issn: 0887-6924
e-issn: 1476-5551
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