Abha Khanna, Aazam Alizadeh, Jun Gu, Feng Jiang, Hua-Zhong Zhang, Arnout Ruifrok, Nancy Caraway, Dennis Johnston, Ruth L Katz. University of Texas, M.D.Anderson Cancer Center, Houston, TX; University of Texas, M. D.Anderson Cancer Center, Houston, TX.

Introduction
Urologic cytology is not sensitive in diagnosing urothelial carcinoma (UC).  Fluorescence-in-situ-hybridization (FISH) for multiple centromeric probes has previously been shown to be a very sensitive test for diagnosing UC, however the test was limited by the requirement of multiple cytospins to evaluate 4 or more probe sets.  Recently a new commercial test (VYSIS) for evaluating urinary cytology became available in which 4 probes are simultaneously evaluated on a per cell basis on a single cytospin.  We performed a pilot study to test the efficacy of the new FISH test compared to standard urine cytology.

Materials and Methods
14 urinary specimens were reacted with a DNA probe set labeled with different fluorochromes and directed against the chromosomes 3,7, and 17, and a locus specific probe against 9p21.  25 cells were scored for each probe.  Normal control bladder wash showed 2 signals (diploid) for each probe.  Abnormal signals were defined as non-diploid.

Results.
11/14 specimens were evaluable.  The number of abnormal cells ranged from 14 to 25 with a median of 22 abnormal cells.  Of 6 cases with 22 or less abnormal cells, 4 were scored as atypical by cytology and 2 as UC.  All 5 cases with >2 abnormal cells were high grade UC.  Two cases of high grade UC showed 90% homozygous deletion for 9p21, while the remaining cases showed a wide range of LOH for 9p21.

Conclusion
This study showed that the multi-color FISH probe test was more sensitive than cytology, easily performed and yielded a high number of cells with numerical chromosomal aberrations.  Further studies are needed to evaluate these results prospectively against follow-up biopsies.

*This abstract has been selected for platform presentation at the 48th Annual Scientific Meeting of the American Society of Cytopathology, Philadelphia,Nov.7-11, 2000.

Reference: The Development of a Multitarget, Multicolor Fluorescence in Situ Hybridization Assay for the Detection of Urothelial Carcinoma in Urine. Irina A et.al. Journal of Molecular Diagnostics, (2)3 116-123, 2000) 
 

Figure 1. Cytology from a normal urine specimen showing normal morphology.	Figure 2. Corresponding diploid DNA histogram from the same specimen shown left .	Figure 3. Normal UroVysion FISH pattern from the same specimen shown left.
Figure 4. Cytology from a low grade UC patient showing apparently normal  morphology.	Figure 5. Corresponding false negative diploid DNA histogram from the same specimen shown left.	Figure 6. Aaneuploid UroVysion FISH pattern from the same specimen shown left.
Figure 7. Cytology from a patient with high grade UC showing abnormal morphology.	Figure 8. Corresponding aneuploid DNA histogram from the same patient  shown left .	Figure 9. Aaneuploid UroVysion FISH pattern from the same patient shown left.