Comparison of a Multiplex Reverse Transcriptasendash;Polymerase Chain Reaction forBCR-ABLto Fluorescence In Situ Hybridization, Southern Blotting, and Conventional Cytogenetics in the Monitoring of Patients WithPh1-Positive Leukemias

摘要

A multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) assay for both major forms ofBCR-ABLwas compared with fluorescence in situ hybridization (FISH), karyotyping, and Southern blotting for disease monitoring in 37 follow-up bone marrow samples from 32 patients withPh1-positive leukemia. Of these 37 samples, 33 were from patients with chronic myeloid leukemia (CML) (26 post allogeneic bone marrow transplantation lsqb;AlloBMTrsqb; and seven during interferon-agr; therapy) and 4 fromPh1-positive acute lymphoblastic leukemia (ALL) patients (1 post AlloBMT and 3 post high dose chemotherapy). For the 27 samples studied after AlloBMT (26 CML and 1Ph1-positive ALL) the time after transplantation ranged from 1 to 107 months (median 47.5 months). In 8 (22percnt;) of the 37 samples there were discrepant results among methods. The discrepancy rates relative to other techniques werecolon; karyotyping 17percnt; (5 of 29), Southern blotting 18percnt; (6 of 33), multiplex RT-PCR 8percnt; (3 of 37), and FISH 8percnt; (3 of 37). Therefore, the relative accuracy of each method for disease monitoring inPh1-positive leukemia wascolon; 83percnt; (24 of 29) for karyotyping, 82percnt; (27 of 33) for Southern blotting, 92percnt; (34 of 37) for FISH, and 92percnt; (34 of 37) for multiplex RT-PCR. This multiplex RT-PCR assay appears equivalent to FISH in terms of accuracy, simplicity, and turnaround time and both are superior to Southern blot and conventional cytogenetics in the laboratory monitoring ofPh1-positive leukemias.