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Vol 50(2016) N 3 p. 412-416; DOI 10.1134/S0026893316020084 Full Text

A.Yu. Ikonnikova1, Yu.E. Yatsenko1, O.S. Kremenetskaya2, O.V. Vinogradova3, D.O. Fesenko1, I.S. Abramov1, V.A. Ovsepyan4, T.V. Nasedkina1*

Detection of BCR-ABL gene mutations in chronic myeloid leukemia using biochips

1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia
2Center for Theoretical Problems of Physicochemical Pharmacology, Russian Academy of Sciences, Moscow, 119991, Russia
3Rogachev Federal Research Clinical Center of Pediatric Hematology, Oncology and Immunology, Ministry of Health of the Russian Federation, Moscow, 117997, Russia
4Kirov Research Institute of Hematology and Blood Transfusion, Kirov, 610027, Russia


*nased@eimb.ru
Received - 2014-11-04; Accepted - 2015-09-29

A biochip-based method was developed to identify the BCR-ABL mutations that affect the thyrosine kinase domain and determine resistance to targeted therapy with thyrosine kinase inhibitors. The method is based on RT-PCR followed by allele-specific hybridization on a biochip with immobilized oligonucleotide probes. The biochip addresses 11 mutations, which are responsible for up to 85% of imatinib resistance cases. A method to decect the clinically significant mutation T315I was designed on the basis of LNA-clamped PCR and proved highly sensitive, detecting the mutation in clinical samples with a leukemic cell content of 5% or higher. The method was validated using clinical samples from chronic myeloid leukemia (CML) patients with acquired resistance to imatinib. The results of hybridization on biochip were verified by Sanger sequencing.

chronic myeloid leukemia, chimeric gene BCR-ABL, mutations, imatinib resistance



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