Rapid and sensitive allele-specific (AS)-RT-PCR assay for detection of T315I mutation in chronic myeloid leukemia patients treated with tyrosine-kinase inhibitors
| Autor: | Roberta Bittencourt, Verónica Pérez, Rosario Uriarte, Arthur Moellmann, Vanesa Scholl, Lem Martinez, Jane Dobbin, Rocio Hassan, Gonzalo Manrique Arechavaleta, Ilana Zalcberg Renault, Héctor N. Seuánez |
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| Rok vydání: | 2010 |
| Předmět: |
medicine.medical_specialty
Drug Resistance Mutation Missense Drug resistance Sensitivity and Specificity General Biochemistry Genetics and Molecular Biology Leukemia Myelogenous Chronic BCR-ABL Positive hemic and lymphatic diseases Internal medicine medicine Humans Missense mutation Enzyme Inhibitors Alleles Hematology Clinical Laboratory Techniques Reverse Transcriptase Polymerase Chain Reaction business.industry Point mutation Reproducibility of Results Myeloid leukemia Imatinib General Medicine Protein-Tyrosine Kinases medicine.disease Molecular biology Leukemia Early Diagnosis Amino Acid Substitution Proto-Oncogene Proteins c-bcr Cancer research business Tyrosine kinase medicine.drug |
| Zdroj: | Clinical and Experimental Medicine. 11:55-59 |
| ISSN: | 1591-9528 1591-8890 |
| DOI: | 10.1007/s10238-010-0101-x |
| Popis: | Point mutations in the kinase domain of BCR-ABL were described in 40-90% of patients with chronic myeloid leukemia (CML) resistant to Imatinib. We herein describe the development of a rapid allele-specific (AS)-RT-PCR assay to identify the T315I mutation, which confers full resistance to all available tyrosine-kinase inhibitors (TKI). The mutation status of 65 patients with resistant CML was evaluated, and the T315I was detected in 3/65 (4.6%). Comparisons between sequencing and AS-RT-PCR results, as well as serial dilutions experiments proved that the method is specific and reproducible, with maximum sensitivity of 1 × 10(-3). The developed assay is a convenient and easy tool to be used in research of CML resistance for rapid mutation screening and, together with sequencing, may be included in efficient strategies for early detection of TKI resistance in patients with CML. |
| Databáze: | OpenAIRE |
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