alexa Utilization of Next-Generation Deep Sequencing to Analy
ISSN: 2329-6917

Journal of Leukemia
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Research Article

Utilization of Next-Generation Deep Sequencing to Analyze BCR-ABL1 Kinase Domain Mutation for Imatinib-Resistant Chronic Myeloid Leukemia Patients in Vietnam

Duong CQ1#, Nguyen C2#, Nguyen TT1, Nguyen LV2, Pham HQ2, Trinh HTT2, Tran HC1, Le TQ1, Pham HT1, Hong TH1, Nguyen TH1, Truong HN2, Bach KQ1 and Nguyen TA1*

1National Institute of Hematology and Blood Transfusion (NIHBT), No. 1, Pham Van Bach street, Cau Giay, Hanoi, Vietnam

2Institute of Biotechnology (IBT), No. 18, Hoang Quoc Viet street, Cau Giay, Hanoi, Vietnam

#The first two authors have contributed equally to this work

*Corresponding Author:
Tri Anh Nguyen
National Institute of Hematology and Blood Transfusion (NIHBT)
No. 1, Pham Van Bach street, Cau GiayHanoi, Vietnam
Tel: +84-4-3782-1985
Fax: +84-4-3868-5582
E-mail: [email protected]

Received date: May 10, 2017; Accepted date: May 27, 2017; Published date: June 05, 2017

Citation: Duong CQ, Nguyen C, Nguyen TT, Nguyen LV, Pham HQ, et al. (2017) Utilization of Next-Generation Deep Sequencing to Analyze BCR-ABL1 Kinase Domain Mutation for Imatinib-Resistant Chronic Myeloid Leukemia Patients in Vietnam. J Leuk 5:235. doi: 10.4172/2329-6917.1000235

Copyright: © 2017 Dupong CQ, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

Background: Chronic myeloid leukemia is a clonal myeloproliferative neoplasm, characterized by the presence of chromosomal translocation t(9; 22)(q34; q11). This is found in over 95% of the cases and results in the BCR-ABL1 fusion protein with high tyrosine kinase activity. During the last decades, imatinib and other generations of tyrosine kinase inhibitor have been used effectively for target therapy of the disease. However, many of the drug resistant cases have been reported recently, due to the mutation within kinase domain of the BCR-ABL1 fusion gene. In order to provide further information about this incidence, we performed a retrospective study of 141 imatinib-resistance chronic myeloid leukemia patients to analyze kinase domain mutation by deep sequencing. Another group of 20 untreated patients were added as control. Methods: RNA from bone marrow cells were extracted and followed by cDNA synthesis. Nested polymerase chain reaction was performed to amplify kinase domain of the BCR-ABL1 fusion gene. The amplified products were monitored size, concentration and prepared DNA sequencing library. Sequence analysis was performed using Illumina MiSeq sequencer and Sequence Pilot software. The sequencing results were randomly chose for Sanger sequencing. Results: None of the control group was positive with kinase domain mutation. There were 47 out of 141 patients (33%) detected with at least one nucleotide substitution. The sequencing results were also confirmed by Sanger sequencing. Of those 47 samples, 72 nucleotide substitutions of 28 types, altered 24 codons were identified. Among those, Y253F/H, M351T, G250E, F359V/I and M244V were the most frequent mutations, while T315I took only 4.1%. There were also a number of samples harboring multiple substitutions and new variations. Conclusion: Nextgeneration deep sequencing is a sensitive and effective method to detect kinase domain mutation and our results could provide further information about the drug-resistance mutation in chronic myeloid leukemia.

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