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Improved Sensitivity of BCR-ABL Detection: A Triple-Probe Three-Color Fluorescence In Situ Hybridization System
P.B. Sinclair,
A.R. Green,
C. Grace, and
E.P. Nacheva
From the Department of Haematology, Addenbrooke's Hospital, Cambridge, UK; the Department of Haematology, University of Cambridge, MCR Center, Cambridge, UK; and Digital Scientific, Cambridge, UK.
Chronic myeloid leukemia is a clonal stem cell disorder associated with the Philadelphia (Ph) translocation [t(9; 22) (q34; q11)]. As a result of the Ph translocation, parts of the ABL and BCR genes become fused. Cytogenetic quantification of Ph+ metaphases can be used to monitor patient response to treatment but is of limited sensitivity and applies only to cycling cells. Fluorescence in situ hybridization (FISH) with probes from the BCR and ABL regions can also identify the Ph translocation in interphase cells. Established systems for the detection of fusion genes by FISH rely on colocalization of two different probes but are associated with a high rate of false-positive results. We have introduced a third probe labeled with a different fluorochrome to create a triple-probe/three-color system that permits identification of both the Ph chromosome and the derivative 9 chromosome in Ph+ cells. This system was used to determine the frequency of interphase cells carrying the BCR-ABL fusion gene in bone marrow and peripheral blood granulocytes from patients showing variable cytogenetic responses to interferon. Our data show that the triple-probe/three-color approach allows highly sensitive detection of residual disease. Moreover, this method is readily applicable to the analysis of other chromosome translocations.
Blood, Vol. 90 No. 4 (August 15), 1997:
pp. 1395-1402
© 1997 by The American Society of Hematology.
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