{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T09:20:26Z","timestamp":1785316826243,"version":"3.55.0"},"reference-count":28,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,4,15]],"date-time":"2025-04-15T00:00:00Z","timestamp":1744675200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Missense mutations in the BCR::ABL1 kinase domain are found in approximately 12\u201380% of patients with chronic myeloid leukemia (CML). Clinically significant mutations include T315I, M244V, Y253H\/F, E255K\/V, V299L, and F359V. The aim of this study was to create a diagnostic system for rapid and inexpensive detection of the above mutations. We used genomic DNA and RNA from peripheral blood and bone marrow cells of 57 patients with a Ph-positive CML diagnosis established in the chronic phase. We have developed a method to detect mutations in the BCR::ABL1 gene based on allele-specific real-time polymerase chain reaction (AS-PCR). In parallel, we analyzed the RNA sequence of the protein kinase domain of the same samples by next-generation sequencing (NGS) covering the points of putative mutations. In this work, we compared the results obtained by both methods for mutation detection and variant allele frequency (VAF) estimation of mutated vs. normal alleles. The sensitivity and specificity of our diagnostic system were also evaluated. It was found that AS-PCR gives reliable results at VAF up to 0.01%. AS-PCR has high sensitivity and may serve as an alternative for the more time-consuming NGS in some cases, as well as for monitoring CML treatment and for analyzing archival material.<\/jats:p>","DOI":"10.3390\/ijms26083728","type":"journal-article","created":{"date-parts":[[2025,4,15]],"date-time":"2025-04-15T06:50:27Z","timestamp":1744699827000},"page":"3728","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Allele-Specific PCR for Detection of Missense Mutations in the Chimeric BCR::ABL1 Gene Causing Failure of Tyrosine Kinase Inhibitor Therapy in CML Patients"],"prefix":"10.3390","volume":"26","author":[{"given":"Anastasia","family":"Skripkina","sequence":"first","affiliation":[{"name":"National Medical Research Center for Hematology, 125167 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irina","family":"Fevraleva","sequence":"additional","affiliation":[{"name":"National Medical Research Center for Hematology, 125167 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Kuzmina","sequence":"additional","affiliation":[{"name":"National Medical Research Center for Hematology, 125167 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6253-3334","authenticated-orcid":false,"given":"Bella","family":"Biderman","sequence":"additional","affiliation":[{"name":"National Medical Research Center for Hematology, 125167 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Stepanova","sequence":"additional","affiliation":[{"name":"National Medical Research Center for Hematology, 125167 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ekaterina","family":"Chelysheva","sequence":"additional","affiliation":[{"name":"National Medical Research Center for Hematology, 125167 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Turkina","sequence":"additional","affiliation":[{"name":"National Medical Research Center for Hematology, 125167 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9463-9187","authenticated-orcid":false,"given":"Andrey","family":"Sudarikov","sequence":"additional","affiliation":[{"name":"National Medical Research Center for Hematology, 125167 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1016\/j.ccell.2020.03.006","article-title":"Response and Resistance to BCR-ABL1-Targeted Therapies","volume":"37","author":"Braun","year":"2020","journal-title":"Cancer Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"670","DOI":"10.3897\/folmed.63.e63366","article-title":"Clinical characteristics, disease evolution and survival in patients with chronic myeloid leukemia, BCR-ABL1 (+) and T315I mutation","volume":"63","author":"Linev","year":"2021","journal-title":"Folia Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/j.clml.2018.06.031","article-title":"Comparison of Frequency and Sensitivity of BCR-ABL1 Kinase Domain Mutations in Asian and White Patients With Imatinib-resistant Chronic-Phase Chronic Myeloid Leukemia","volume":"18","author":"Kim","year":"2018","journal-title":"Clin. 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