Four-channel asymmetric Real-Time PCR hybridization probe assay: A rapid pre-screening method for critical BCR-ABL kinase domain mutations
| Autor: | Jordi Martinez-Serra, Carmen Ballester, Carmen Vidal, Josep Miquel Bauça, Sara SanFelix, Joan Besalduch, Teresa Bex, Juan Carlos Amat, Simona Soverini, Carmen Santos, Toni F. Marcús, Andrés Novo, Antonio Gutierrez, María Navarro-Palou, Teresa Ros |
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| Přispěvatelé: | Martinez-Serra J., Gutiérrez A., Marcús T.F., Soverini S., Amat J.C., Navarro-Palou M., Ros T., Bex T., Ballester C., Bauça J.M., Sanfelix S., Novo A., Vidal C., Santos C., Besalduch J. |
| Rok vydání: | 2012 |
| Předmět: |
Male
Asymmetric NILOTINIB DNA Mutational Analysis Clinical Biochemistry Fusion Proteins bcr-abl Nucleic Acid Denaturation Piperazines chemistry.chemical_compound Bone Marrow hemic and lymphatic diseases Fluorescence Resonance Energy Transfer ponatinib BCR-ABL Aged 80 and over DASATINIB ABL Hybridization probe Ponatinib Myeloid leukemia General Medicine Middle Aged Dasatinib Real-time polymerase chain reaction Benzamides Imatinib Mesylate Female DNA Probes medicine.drug Adult kinase Antineoplastic Agents Biology Real-Time Polymerase Chain Reaction IMATINIB Leukemia Myelogenous Chronic BCR-ABL Positive medicine Humans Aged Retrospective Studies Base Sequence Imatinib Molecular biology Pyrimidines chemistry Nilotinib Drug Resistance Neoplasm Mutation FRET real-time PCR Multiplex Polymerase Chain Reaction |
| Zdroj: | Clinical Biochemistry. 45:345-351 |
| ISSN: | 0009-9120 |
| DOI: | 10.1016/j.clinbiochem.2011.12.026 |
| Popis: | Objectives Within the laboratory protocols, used for the study of BCR-ABL resistance mutations in chronic myeloid leukemia patients treated with Imatinib, direct sequencing remains the reference method. Since the incidence of patients with a mutation-related loss of response is not very high, it is very useful in the routine laboratory to perform a fast pre-screening method. Design and methods With this in mind, we have designed a new technique, based on a single Real-Time FRET-based PCR, followed by a study of melting peaks. This new tool, developed in a LightCycler 2.0, combines four different fluorescence channels for the simultaneous detection, in a single close tube, of critical mutations within the ABL kinase domain. Results Assay evaluation performed on 33 samples, previously genotyped by sequentiation, resulted in full concordance of results. Conclusions This new methodology detects in a few steps the presence of critical mutations associated to Imatinib resistance. |
| Databáze: | OpenAIRE |
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