title: Flow Cytometric Immunobead Assay for Detection of BCR-ABL1 Fusion Proteins in Chronic Myleoid Leukemia: Comparison with FISH and PCR Techniques.
abstract: Chronic Myeloid Leukemia (CML) is characterized by a balanced translocation juxtaposing the Abelson (ABL) and breakpoint cluster region (BCR) genes. The resulting BCR-ABL1 oncogene leads to increased proliferation and survival of leukemic cells. Successful treatment of CML has been accompanied by steady improvements in our capacity to accurately and sensitively monitor therapy response. Currently, measurement of BCR-ABL1 mRNA transcript levels by real-time quantitative PCR (RQ-PCR) defines critical response endpoints. An antibody-based technique for BCR-ABL1 protein recognition could be an attractive alternative to RQ-PCR. To date, there have been no studies evaluating whether flow-cytometry based assays could be of clinical utility in evaluating residual disease in CML patients. Here we describe a flow-cytometry assay that detects the presence of BCR-ABL1 fusion proteins in CML lysates to determine the applicability, reliability, and specificity of this method for both diagnosis and monitoring of CML patients for initial response to therapy. We show that: i) CML can be properly diagnosed at onset, (ii) follow-up assessments show detectable fusion protein (i.e. relative mean fluorescent intensity, rMFI%>1) when BCR-ABL1IS transcripts are between 1-10%, and (iii) rMFI% levels predict CCyR as defined by FISH analysis. Overall, the FCBA assay is a rapid technique, fully translatable to the routine management of CML patients.
authors.affiliation: Biotechnology Research Unit, ASP Cosenza, Aprigliano, Italy.
authors.forename: Anna Grazia
authors.initials: AG
authors.lastname: Recchia
authors.affiliation: Hematology Unit, Azienda Ospedaliera Annunziata di Cosenza, Cosenza, Italy.
authors.forename: Nadia
authors.initials: N
authors.lastname: Caruso
authors.affiliation: Biotechnology Research Unit, ASP Cosenza, Aprigliano, Italy.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Bossio
authors.affiliation: Biotechnology Research Unit, ASP Cosenza, Aprigliano, Italy.
authors.forename: Mariavaleria
authors.initials: M
authors.lastname: Pellicanò
authors.affiliation: Biotechnology Research Unit, ASP Cosenza, Aprigliano, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: De Stefano
authors.affiliation: Hematology Unit, Azienda Ospedaliera Annunziata di Cosenza, Cosenza, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Franzese
authors.affiliation: Biotechnology Research Unit, ASP Cosenza, Aprigliano, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Palummo
authors.affiliation: Department of Oncology, Hematology and Bone Marrow Transplantation Unit, University of Palermo, Policlinico P. Giaccone, Palermo, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Abbadessa
authors.affiliation: Hematology Unit, Azienda Ospedaliera Annunziata di Cosenza, Cosenza, Italy.
authors.forename: Eugenio
authors.initials: E
authors.lastname: Lucia
authors.affiliation: Hematology Unit, Azienda Ospedaliera Annunziata di Cosenza, Cosenza, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Gentile
authors.affiliation: Hematology Unit, Azienda Ospedaliera Annunziata di Cosenza, Cosenza, Italy.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Vigna
authors.affiliation: Department of Oncology, Hematology and Bone Marrow Transplantation Unit, University of Palermo, Policlinico P. Giaccone, Palermo, Italy.
authors.forename: Clementina
authors.initials: C
authors.lastname: Caracciolo
authors.affiliation: Centro Trasfusionale Ospedale, Azienda Sanitaria Provinciale 7, Ragusa, Italy.
authors.forename: Antolino
authors.initials: A
authors.lastname: Agostino
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Galimberti
authors.affiliation: Medical Oncology Unit, Hematology-Oncology Department, Azienda Ospedaliera Pugliese-Ciaccio, Catanzaro, Italy.
authors.forename: Luciano
authors.initials: L
authors.lastname: Levato
authors.affiliation: Divisione di Ematologia, Ospedale Ferrarotto, Università degli Studi di Catania, Catania, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Stagno
authors.affiliation: Medical Oncology Unit, Hematology-Oncology Department, Azienda Ospedaliera Pugliese-Ciaccio, Catanzaro, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Molica
authors.affiliation: U.O.C. di Ematologia dell'Azienda"Bianchi-Melacrino-Morelli" di Reggio Calabria, Reggio Calabria, Italy.
authors.forename: Bruno
authors.initials: B
authors.lastname: Martino
authors.affiliation: Dipartimento di Scienze Mediche e Pediatriche, Università degli Studi di Catania, Catania, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Vigneri
authors.affiliation: Divisione di Ematologia, Ospedale Ferrarotto, Università degli Studi di Catania, Catania, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Di Raimondo
authors.affiliation: Biotechnology Research Unit, ASP Cosenza, Aprigliano, Italy; Hematology Unit, Azienda Ospedaliera Annunziata di Cosenza, Cosenza, Italy.
authors.forename: Fortunato
authors.initials: F
authors.lastname: Morabito
mesh_terms: D005434:Flow Cytometry; D016044:Fusion Proteins, bcr-abl / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007118:Immunoassay; D017404:In Situ Hybridization, Fluorescence; D015477:Leukemia, Myelomonocytic, Chronic / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D016133:Polymerase Chain Reaction
chemical_list: C581634:BCR-ABL1 fusion protein, human; D016044:Fusion Proteins, bcr-abl