Publication: A single-tube allele specific-polymerase chain reaction to detect T315I resistant mutation in chronic myeloid leukemia patients
Issued Date
2011-02-10
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ISSN
17568722
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2-s2.0-79551688229
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Hematology and Oncology. Vol.4, (2011)
Suggested Citation
Wanwisa Wongboonma, Wanna Thongnoppakhun, Chirayu U. Auewarakul A single-tube allele specific-polymerase chain reaction to detect T315I resistant mutation in chronic myeloid leukemia patients. Journal of Hematology and Oncology. Vol.4, (2011). doi:10.1186/1756-8722-4-7 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/11590
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Title
A single-tube allele specific-polymerase chain reaction to detect T315I resistant mutation in chronic myeloid leukemia patients
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Abstract
Background: BCR-ABL kinase domain (KD) mutation is the major mechanism contributing to suboptimal response to tyrosine kinase inhibitors (TKI) in BCR-ABL-positive chronic myeloid leukemia (CML) patients. T315I mutation, as one of the most frequent KD mutations, has been shown to be strongly associated with TKI resistance and subsequent therapeutic failure. A simple and sensitive method is thus required to detect T315I mutation at the earliest stage. Methods. A single-tube allele specific-polymerase chain reaction (AS-PCR) method was developed to detect T315I mutation in a mixture of normal and mutant alleles of varying dilutions. Denaturing high performance liquid chromatography (DHPLC) and direct sequencing were performed as a comparison to AS-PCR. Results: T315I mutant bands were observed in the mixtures containing as low as 0.5-1% of mutant alleles by AS-PCR. The detection sensitivity of DHPLC was around 1.5-3% dilution whereas sequencing analysis was unable to detect below 6.25% dilution. Conclusion: A single-tube AS-PCR is a rapid and sensitive screening method for T315I mutation. Detection of the most resistant leukemic clone in CML patients undergoing TKI therapy should be feasible with this simple and inexpensive method. © 2011 Wongboonma et al; licensee BioMed Central Ltd.