Rapid and sensitive allele-specific (AS)-RT-PCR assay for detection of T315I mutation in chronic myeloid leukemia patients treated with tyrosine-kinase inhibitors

被引：9

作者：

Manrique Arechavaleta, Gonzalo ^{[1
]}

Scholl, Vanesa ^{[2
]}

Perez, Veronica ^{[1
]}

Bittencourt, Roberta ^{[2
]}

Moellmann, Arthur ^{[3
]}

Hassan, Rocio ^{[2
]}

Seuanez, Hector N. ^{[4
]}

Dobbin, Jane ^{[3
]}

Martinez, Lem ^{[1
]}

Renault, Ilana Zalcberg ^{[2
]}

Uriarte, Rosario ^{[1
]}

机构：

[1] Asociac Espanola Primera Socorros Mutuos, Div Mol Biol, Specialized Tech Lab, Montevideo 11200, Uruguay

[2] INCA, Bone Marrow Transplantat Ctr CEMO, Mol Biol Lab, Rio De Janeiro, Brazil

[3] INCA, Hematol Serv, Rio De Janeiro, Brazil

[4] INCA, Div Genet, Rio De Janeiro, Brazil

来源：

CLINICAL AND EXPERIMENTAL MEDICINE | 2011年 / 11卷 / 01期

关键词：

Chronic myeloid leukemia; BCR-ABL; Kinase domain mutations; AS-RT-PCR; BCR-ABL TRANSCRIPTS; IDENTIFY PATIENTS; DOMAIN MUTATIONS; IMATINIB; CML; PCR; RESISTANCE;

D O I：

10.1007/s10238-010-0101-x

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

1001 ;

摘要：

Point mutations in the kinase domain of BCR-ABL 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 x 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.

引用

页码：55 / 59

页数：5

共 14 条

[1] Evolving concepts in the management of chronic myeloid leukemia: recommendations from an expert panel on behalf of the European LeukemiaNet [J].

Baccarani, Michele ;

Saglio, Giuseppe ;

Goldman, John ;

Hochhaus, Andreas ;

Simonsson, Bengt ;

Appelbaum, Frederick ;

Apperley, Jane ;

Cervantes, Francisco ;

Cortes, Jorge ;

Deininger, Michael ;

Gratwohl, Alois ;

Guilhot, Frangois ;

Horowitz, Mary ;

Hughes, Timothy ;

Kantarjian, Hagop ;

Larson, Richard ;

Niederwieser, Dielger ;

Silver, Richard ;

Hehlmann, Rudiger .

BLOOD, 2006, 108 (06) :1809-1820

[2] High frequency of point mutations clustered within the adenosine triphosphate-binding region of BCR/ABL in patients with chronic myeloid leukemia or Ph-positive acute lymphoblastic leukemia who develop imatinib (STI571) resistance [J].

Branford, S ;

Rudzki, Z ;

Walsh, S ;

Grigg, A ;

Arthur, C ;

Taylor, K ;

Herrmann, R ;

Lynch, KP ;

Hughes, TP .

BLOOD, 2002, 99 (09) :3472-3475

[3] Detection of BCR-ABL mutations in patients with CML treated with imatinib is virtually always accompanied by clinical resistance, and mutations in the ATP phosphate-binding loop (P-loop) are associated with a poor prognosis [J].

Branford, S ;

Rudzki, Z ;

Walsh, S ;

Parkinson, I ;

Grigg, A ;

Szer, J ;

Taylor, K ;

Herrmann, R ;

Seymour, JF ;

Arthur, C ;

Joske, D ;

Lynch, K ;

Hughes, T .

BLOOD, 2003, 102 (01) :276-283

[4] Real-time quantitative PCR analysis can be used as a primary screen to identify patients with CML treated with imatinib who have BCR-ABL kinase domain mutations [J].

Branford, S ;

Rudzki, Z ;

Parkinson, I ;

Grigg, A ;

Taylor, K ;

Seymour, JF ;

Durrant, S ;

Browett, P ;

Schwarer, AP ;

Arthur, C ;

Catalano, J ;

Leahy, MF ;

Filshie, R ;

Bradstock, K ;

Herrmann, R ;

Joske, D ;

Lynch, K ;

Hughes, T .

BLOOD, 2004, 104 (09) :2926-2932

[5]

Branford Susan, 2007, Hematology Am Soc Hematol Educ Program, P376

[6]

Clayton SJ, 2000, CLIN CHEM, V46, P1929

[7] Monitoring the response and course of chronic myeloid leukemia in the modern era of BCR-ABL tyrosine kinase inhibitors: practical advice on the use and interpretation of monitoring methods [J].

Kantarjian, Hagop ;

Schiffer, Charles ;

Jones, Dan ;

Cortes, Jorge .

BLOOD, 2008, 111 (04) :1774-1780

[8] European LeukemiaNet criteria for failure or suboptimal response reliably identify patients with CML in early chronic phase treated with imatinib whose eventual outcome is poor [J].

Marin, David ;

Milojkovic, Dragana ;

Olavarria, Eduardo ;

Khorashad, Jamshid S. ;

de lavallade, Hugues ;

Reid, Alistair G. ;

Foroni, Letizia ;

Rezvani, Katayoun ;

Bua, Marco ;

Dazzi, Francesco ;

Pavlu, Jiri ;

Klammer, Matthias ;

Kaeda, Jaspal S. ;

Goldman, John M. ;

Apperley, Jane F. .

BLOOD, 2008, 112 (12) :4437-4444

[9] State-of-the-art in the treatment of chronic myeloid leukaemia [J].

Milojkovic, Dragana ;

Apperley, Jane .

CURRENT OPINION IN ONCOLOGY, 2008, 20 (01) :112-121

[10] Bcr-Abl kinase domain mutations, drug resistance, and the road to a cure for chronic myeloid leukemia [J].

O'Hare, Thomas ;

Eide, Christopher A. ;

Deininger, Michael W. N. .

BLOOD, 2007, 110 (07) :2242-2249

← 1 2 →