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Detection of chimeric BCR-ABL genes on bone marrow samples and blood smears
in chronic myeloid and acute lymphoblastic leukemia by in situ
hybridization
M Bentz, G Cabot, M Moos, MR Speicher, A Ganser, P Lichter and H Dohner
Medizinische Klinik, Universitat Heidelberg, Germany.
The presence of BCR-ABL fusion genes has important diagnostic and
prognostic implications in chronic myeloid leukemia (CML) and acute
lymphoblastic leukemia (ALL). The CML-specific chimeric BCR-ABL gene with a
break involving the major breakpoint cluster region (M-bcr) of the BCR-gene
has been detected by means of fluorescence in situ hybridization (FISH). In
this study, we present a FISH protocol that allows the detection of breaks
in both the major and the minor breakpoint cluster region (m-bcr). Three
hybridization signals of D107F9, a yeast artificial chromosome
(YAC)-derived probe spanning the breakpoint regions of the BCR gene, were
indicative of the translocation events. To increase the specificity
further, this probe was combined with cos-abl 8, a cosmid probe flanking
the breakpoint within the ABL gene for dual-color hybridization. Samples of
21 patients with CML, the ALL-derived cell line SUP-B15, and of seven
patients with Philadelphia chromosome (Ph1)-positive ALL (three of them
with breakpoints within m-bcr) were examined. BCR-ABL fusion was detected
in all cases with high specificity (false-positive nuclei: mean, 0.1%). On
cytogenetic preparations, the percentages of BCR-ABL- positive interphase
cells ranged from 53% to 91%. Comparable efficiencies were achieved on
blood smears. In conclusion, hybridization with D107F9 and cos-abl 8 allows
unambiguous diagnosis of BCR-ABL genes and is likely to become an important
tool for the monitoring of therapies in patients with CML and ALL.
Volume 83,
Issue 7,
pp. 1922-1928,
04/01/1994
Copyright © 1994 by The American Society of Hematology
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