FIELD: medicine.
SUBSTANCE: invention refers to medicine, particularly to medical genetics and oncohematology, and aims at quantitative determination of mutations F317L and F359V/C of the BCR-ABL kinase domain in patients suffering chronic myeloid leukemia resistant to tyrosine kinase inhibitors. One stage of PCR is used to determine expression of BCR-ABLF317L and BCR-ABLF359V/C. Primers used are a system for determining F317L consisting of a forward primer SEQ ID NO: 1 and reverse specific primers for mutation SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 and specific probe SEQ ID NO: 5. Determining expression of F359V/C, BCR-ABL and ABL, using systems consisting of primers and probes. Number of copies of BCR-ABLF317L and BCR-ABLF359V/C in samples are determined by calibration curve, which is constructed according to standards with known concentration. BCR-ABLF317L, BCR-ABLF359V/C and BCR-ABL expression is calculated relative to control ABL gene by formulas.
EFFECT: invention provides an effective method of determining mutations F317L and F359V/C of the BCR-ABL kinase domain, having higher sensitivity and specificity.
1 cl, 1 dwg
Authors
Dates
2019-08-27—Published
2019-03-26—Filed