TY - JOUR
T1 - Rapid screening of leukemia fusion transcripts in acute leukemia by real-time PCR
AU - Osumi, Kazuoki
AU - Fukui, Takafumi
AU - Kiyoi, Hitoshi
AU - Kasai, Masanobu
AU - Kodera, Yoshihisa
AU - Kudo, Kazuko
AU - Kato, Koji
AU - Matsuyama, Takaharu
AU - Naito, Kazuyuki
AU - Tanimoto, Mitsune
AU - Hirai, Hisamaru
AU - Saito, Hidehiko
AU - Ohno, Ryuzo
AU - Naoe, Tomoki
PY - 2002/12/1
Y1 - 2002/12/1
N2 - We established a real-time PCR method that can simultaneously detect 10 different fusion transcripts (major, minor and micro BCR/ABL, AML1/MTG8, PML/RARα, CBFβ/MYH11, TEL/AML1, E2A/PBX1, MLL/AF4, and MLL/AF9) together with Wilms' tumor gene (WT1) transcripts. This screening method allowed the processing of six specimens concomitantly and required only one working day from RNA extraction to final results. Fifty-seven bone marrow (BM) samples from patients with acute leukemia were retrospectively screened for the presence of fusion and WT1 transcripts without knowledge of the cytogenetic data, and the fusion transcripts were detected in 20 of 57 samples (35.1%). The concordance between the present method and cytogenetic analysis was examined in 38 samples in which the cytogenetic data were available. In 12 of 38 samples, the PCR results agreed with the cytogenetic data, whereas in 4 of the remaining 26 samples, the translocations were detected by real-time PCR alone because of the insufficient number of metaphases obtained and presumably the submicroscopic or masked translocations. The WT1 levels ranged from 400 to 690,000 copies/μg RNA in BM from leukemia patients, whereas 0-470 copies/μg RNA were found in BM cells from BMT donors. This real-time PCR method enables rapid and efficient characterization of acute leukemia in addition to subsequent evaluation of minimal residual diseases.
AB - We established a real-time PCR method that can simultaneously detect 10 different fusion transcripts (major, minor and micro BCR/ABL, AML1/MTG8, PML/RARα, CBFβ/MYH11, TEL/AML1, E2A/PBX1, MLL/AF4, and MLL/AF9) together with Wilms' tumor gene (WT1) transcripts. This screening method allowed the processing of six specimens concomitantly and required only one working day from RNA extraction to final results. Fifty-seven bone marrow (BM) samples from patients with acute leukemia were retrospectively screened for the presence of fusion and WT1 transcripts without knowledge of the cytogenetic data, and the fusion transcripts were detected in 20 of 57 samples (35.1%). The concordance between the present method and cytogenetic analysis was examined in 38 samples in which the cytogenetic data were available. In 12 of 38 samples, the PCR results agreed with the cytogenetic data, whereas in 4 of the remaining 26 samples, the translocations were detected by real-time PCR alone because of the insufficient number of metaphases obtained and presumably the submicroscopic or masked translocations. The WT1 levels ranged from 400 to 690,000 copies/μg RNA in BM from leukemia patients, whereas 0-470 copies/μg RNA were found in BM cells from BMT donors. This real-time PCR method enables rapid and efficient characterization of acute leukemia in addition to subsequent evaluation of minimal residual diseases.
UR - http://www.scopus.com/inward/record.url?scp=0036890819&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036890819&partnerID=8YFLogxK
U2 - 10.1080/1042819021000040206
DO - 10.1080/1042819021000040206
M3 - Article
C2 - 12613515
AN - SCOPUS:0036890819
SN - 1042-8194
VL - 43
SP - 2291
EP - 2299
JO - Leukemia and Lymphoma
JF - Leukemia and Lymphoma
IS - 12
ER -
