title: Rapid screening of leukemia fusion transcripts in acute leukemia by real-time PCR.
abstract: We established a real-time PCR method that can simultaneously detect 10 different fusion transcripts (major, minor and micro BCR/ABL, AML1/MTG8, PML/RARalpha, CBFbeta/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/microg RNA in BM from leukemia patients, whereas 0-470 copies/microg 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.
authors.affiliation: Otsuka Assay Laboratories, Otsuka Pharmaceutical Co. Ltd., Tokyo, Japan.
authors.forename: Kazuoki
authors.initials: K
authors.lastname: Osumi
authors.forename: Takafumi
authors.initials: T
authors.lastname: Fukui
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kiyoi
authors.forename: Masanobu
authors.initials: M
authors.lastname: Kasai
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Kodera
authors.forename: Kazuko
authors.initials: K
authors.lastname: Kudo
authors.forename: Koji
authors.initials: K
authors.lastname: Kato
authors.forename: Takaharu
authors.initials: T
authors.lastname: Matsuyama
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Naito
authors.forename: Mitsune
authors.initials: M
authors.lastname: Tanimoto
authors.forename: Hisamaru
authors.initials: H
authors.lastname: Hirai
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Saito
authors.forename: Ryuzo
authors.initials: R
authors.lastname: Ohno
authors.forename: Tomoki
authors.initials: T
authors.lastname: Naoe
mesh_terms: D000208:Acute Disease; D001853:Bone Marrow; D016022:Case-Control Studies; D017931:DNA Primers; D005260:Female; D006801:Humans; D007938:Leukemia / Q000175:diagnosis* / Q000235:genetics; D008297:Male; D018365:Neoplasm, Residual / Q000175:diagnosis; D015514:Oncogene Proteins, Fusion / Q000032:analysis* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D012334:RNA, Neoplasm / Q000032:analysis; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity; D014178:Translocation, Genetic; D009396:Wilms Tumor / Q000175:diagnosis / Q000235:genetics
chemical_list: D017931:DNA Primers; D015514:Oncogene Proteins, Fusion; D012333:RNA, Messenger; D012334:RNA, Neoplasm