Flow Cytometric Immunobead Assay for Detection of BCR-ABL1 Fusion Proteins in Chronic Myleoid Leukemia: Comparison with FISH and PCR Techniques
| Autor: | Ernesto Vigna, Sara Galimberti, Angela Palummo, Fortunato Morabito, Massimo Gentile, Clementina Caracciolo, Luciano Levato, Paolo Vigneri, Stefano Molica, Eugenio Lucia, Nadia Caruso, Anna Grazia Recchia, Bruno Martino, Vincenzo Abbadessa, Antolino Agostino, Francesco Di Raimondo, Stefania Franzese, Mariavaleria Pellicanò, Sabrina Bossio, Fabio Stagno, Laura De Stefano |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Genetics and Molecular Biology (all)
medicine.medical_specialty Science Fusion Proteins bcr-abl Biology Biochemistry Polymerase Chain Reaction Internal medicine hemic and lymphatic diseases medicine Humans Agricultural and Biological Sciences (all) Biochemistry Genetics and Molecular Biology (all) Medicine (all) In Situ Hybridization Fluorescence Immunoassay Multidisciplinary ABL Hematology medicine.diagnostic_test breakpoint cluster region Myeloid leukemia Leukemia Myelomonocytic Chronic medicine.disease Flow Cytometry Molecular biology Fusion protein Leukemia Real-time polymerase chain reaction Medicine Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 10, Iss 6, p e0130360 (2015) |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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