Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: A single-tube allele specific-polymerase chain reaction to detect T315I resistant mutation in chronic myeloid leukemia patients. | Wongboonma et al. Journal of Hematology Oncology 2011 4 7 http content 4 1 7 JOURNAL OF HEMATOLOGY ONCOLOGY RESEARCH Open Access A single-tube allele specific-polymerase chain reaction to detect T315I resistant mutation in chronic myeloid leukemia patients Wanwisa Wongboonma1 Wanna Thongnoppakhun2 Chirayu U Auewarakul3 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 of mutant alleles by AS-PCR. The detection sensitivity of DHPLC was around dilution whereas sequencing analysis was unable to detect below 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. 1. Background Chronic myeloid leukemia CML is a chronic hematopoietic stem cell disorder characterized by extensive proliferation and expansion of myeloid cells at varying stages of maturation and differentiation 1 . The hallmark of CML is the Philadelphia Ph chromosome which occurs as a result of a reciprocal chromosomal translocation between chromosomes 9 and 22 creating a new fusion gene BCR-ABL with constitutive .
