Translocation(15;17) leading to the formation of fusion gene PML/RARalpha is the diagnostic hallmark of acute promyelocytic leukemia (APL). Interphase fluorescence in situ hybridization (FISH) is one of the diagnostic tools employed for the detection of PML/RARalpha rearrangement. Using a dual color dual fusion (D-FISH) PML/RARalpha translocation DNA probe which hybridises both to PML/RARalpha and RARalpha/PML fusion genes, we characterised the FISH pattern of 52 APL patients at diagnosis and correlated the findings with conventional cytogenetics and RT-PCR analysis. The diagnostic sensitivity of the probe for PML/RARalpha was 100%. Seven patients had atypical D-FISH patterns; two had a masked PML/RARalpha fusion signal caused by the insertion of PML into RARalpha on 17q; 3 had an extra copy of PML/RARalpha in the form of isochromosome der(17)(q10)t(15;17) and one had duplication of the normal RARalpha gene with an ider(17q) masquerading as i(17)(q10). There was also one case of t(7;17;15) with a typical D-FISH pattern and in which metaphase FISH suggested an unusual 4-point break. In summary, PML/RARalpha D-FISH is a highly sensitive method for confirming diagnosis of APL. However D-FISH cannot be solely relied on for the diagnosis of APL owing to atypical patterns which are infrequently observed in cases with additional 17q structural abnormalities, gene insertion and gene duplication.