Quantification of minimal residual disease in patients with e1a2 BCR-ABL-positive acute lymphoblastic leukemia using a real-time RT-PCR assay.
Using a real-time RT-PCR method, we analyzed the expression of e1a2 BCR-ABL mRNA in bone marrow samples from 13 patients with e1a2 BCR-ABL-positive acute lymphoblastic leukemia (ALL) at different time points during chemotherapy and after bone marrow transplantation (BMT). The detection limit of the method, assessed using serial dilutions of ALL/MIK cells, was found to be 1:10(5), similar to what is observed for the conventional RT-nested PCR method. The e1a2 BCR-ABL values were normalized with respect to those of the housekeeping gene GAPDH. The decrease in the e1a2 BCR-ABL/GAPDH ratio after remission induction chemotherapy reflects well the response to chemotherapy and consequently correlates with the prognosis. Although molecular remission was achieved by chemotherapy alone, some patients relapsed, and the e1a2 BCR-ABL/GAPDH ratios in these cases progressively increased to the levels seen prior to hematological relapse. Long-term hematological complete remission (more than 30 months) could be achieved in cases in which a more than 4.0 log decrease in the e1a2 BCR-ABL/GAPDH ratio was obtained by chemotherapy alone, and BMT was then performed. In conclusion, real-time RT-PCR allows for an evaluation of the kinetics of e1a2 BCR-ABL/GAPDH expression during the various phases of chemotherapy or after BMT and may be effective for the indication and control of disease relapse in Ph-positive ALL patients.
Yokota
H
H
Department of Laboratory Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Tsuno
N H
NH
Tanaka
Y
Y
Fukui
T
T
Kitamura
K
K
Hirai
H
H
Osumi
K
K
Itou
N
N
Satoh
H
H
Okabe
M
M
Nakahara
K
K
D000328:Adult; D000368:Aged; D016026:Bone Marrow Transplantation; D003131:Combined Modality Therapy; D005260:Female; D016044:Fusion Proteins, bcr-abl / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics; D018922:HL-60 Cells; D006801:Humans; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D018365:Neoplasm, Residual; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis / Q000235:genetics / Q000628:therapy*; D012334:RNA, Neoplasm / Q000096:biosynthesis; D012015:Reference Standards; D012074:Remission Induction; D015203:Reproducibility of Results; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards; D012680:Sensitivity and Specificity; D013997:Time Factors; D016896:Treatment Outcome; D014407:Tumor Cells, Cultured
D012334:RNA, Neoplasm; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D016044:Fusion Proteins, bcr-abl