Rapid and sensitive allele-specific (AS)-RT-PCR assay for detection of T315I mutation in chronic myeloid leukemia patients treated with tyrosine-kinase inhibitors
- نوع فایل : کتاب
- زبان : انگلیسی
- مؤلف : Gonzalo Manrique Arechavaleta Vanesa Scholl Vero´nica Pe´rez Roberta Bittencourt Arthur Moellmann Rocio Hassan He´ctor N. Seua´nez Jane D
- چاپ و سال / کشور: 2010
Description
Point mutations in the kinase domain of BCRABL were described in 40–90% of patients with chronic myeloid leukemia (CML) resistant to Imatinib. We herein describe the development of a rapid allele-specific (AS)- RT-PCR assay to identify the T315I mutation, which confers full resistance to all available tyrosine-kinase inhibitors (TKI). The mutation status of 65 patients with resistant CML was evaluated, and the T315I was detected in 3/65 (4.6%). Comparisons between sequencing and AS-RT-PCR results, as well as serial dilutions experiments proved that the method is specific and reproducible, with maximum sensitivity of 1 9 10-3. The developed assay is a convenient and easy tool to be used in research of CML resistance for rapid mutation screening and, together with sequencing, may be included in efficient strategies for early detection of TKI resistance in patients with CML.
Clin Exp Med (2011) 11:55–59 DOI 10.1007/s10238-010-0101-x Received: 13 October 2009 / Accepted: 23 April 2010 / Published online: 29 May 2010
