Publication: A single-tube allele specific-polymerase chain reaction to detect T315I resistant mutation in chronic myeloid leukemia patients
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Issued Date
2011
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eng
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Mahidol University
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BioMed Central
Bibliographic Citation
Journal of Hematology & Oncology. Vol. 4, (2011), 7
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 & Oncology. Vol. 4, (2011), 7. doi:10.1186/1756-8722-4-7 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/2644
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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.