UroVysion Bladder Cancer Kit

Benefits of Urovysion

• Only urine-based molecular test approved by the FDA to aid bladder cancer surveillance and assist with diagnosis
• Clinically proven by more than 10 years of scientific data
• Greater sensitivity than cytology across all stages and grades
• 100% sensitivity in CIS tumors
• Delivers definitive results in atypical cytology cases with equivocal or negative cystoscopy
• Performs without interference in the presence of substances in urine samples such as BCG, Mitomycin C, and Thiotepa.
• Delivers a 94.1% negative predictive value for detection of bladder cancer recurrence versus cystoscopy/histology in BCG-treated patients

Results from the UroVysion Kit are intended for use, in conjunction with and not in lieu of current standard diagnostic procedures, as an aid for initial diagnosis of bladder carcinoma in patients with hematuria and subsequent monitoring for tumor recurrence in patients previously diagnosed with bladder cancer.

Urovysion Kit Contents

The UroVysion Bladder Cancer Kit probes are directly labeled with one of the Vysis fluorophores; SpectrumRed, SpectrumGreen, SpectrumAqua or SpectrumGold. 

The UroVysion Bladder Cancer Kit consists of:

• Three alpha-satellite repeat sequence probes; CEP 3 SpectrumRed, CEP 7 SpectrumGreen, and CEP 17 SpectrumAqua that hybridize to the centromere regions of chromosomes 3, 7, and 17, respectively. In addition, a unique sequence probe, LSI p16 (9p21) SpectrumGold, is included that hybridizes to the p16 gene at 9p21. 
• This probe set is premixed in hybridization buffer.

Indications and Limitations Of Use

Intended Use

The UroVysion Bladder Cancer Kit (UroVysion Kit) is designed to detect aneuploidy for chromosomes 3, 7, 17, and loss of the 9p21 locus via fluorescence in situ hybridization (FISH) in urine specimens from persons with hematuria suspected of having bladder cancer. Results from the UroVysion Kit are intended for use, in conjunction with and not in lieu of current standard diagnostic procedures, as an aid for initial diagnosis of bladder carcinoma in patients with hematuria and subsequent monitoring for tumor recurrence in patients previously diagnosed with bladder cancer.

Limitations

1. The UroVysion Kit has been optimized for identifying and quantitating chromosomes 3, 7, and 17, and locus 9p21 in human urine specimens.
2. The performance of the UroVysion Kit was validated using the procedures provided in this package insert only. Modifications to these procedures may alter the performance of the assay. 
3. The clinical interpretation of any test results should be evaluated within the context of the patient's medical history and other diagnostic laboratory test results. 
4. UroVysion assay results may not be informative if the specimen quality and/or specimen slide preparation is inadequate, e.g., the presence of excessive granulocytes or massive bacteruria. 
5. Technologists performing the UroVysion signal enumeration must be capable of visually distinguishing between the red and green signals.
6. Positive UroVysion results in the absence of other signs or symptoms of bladder cancer recurrence may be evidence of other urinary tract related cancers, e.g., ureter, urethra, renal, and/or prostate in males, and further patient follow-up is justified. In a study conducted on patients with hematuria (see "Symptomatic Patients: Performance vs. Standard of Care" for details on this clinical study) 3 patients, whose initial bladder cystoscopy was negative, were subsequently diagnosed with renal cancer within 6 months of this initial study visit. All 3 of these cases were positive by UroVysion. 
7. If UroVysion results are negative but standard clinical or diagnostic tests (e.g., cytology, cystoscopy) are positive, the standard procedures take precedence over the UroVysion test. Although the UroVysion Kit was designed to detect genetic changes associated with most bladder cancers, there will be some bladder cancers whose genetic changes cannot be detected by the UroVysion test. 
8. Ta stage solitary tumors smaller than 5mm could not be detected by UroVysion FISH.23 UroVysion FISH results are dependent on the amount of tumor cells that are deposited on the slide.

CAUTION: United States Federal law restricts this device to sale and distribution to or on the order of a physician or to a clinical laboratory; use is restricted to, by, or on the order of a physician.

References

Halling KC, King W, Sokolova A, et al. Assessing the value of reflex fluorescence in situ hybridization testing in the diagnosis of bladder cancer when routine urine cytological examination is equivocal. J Urol. 2000; 164: 1768-1775.

Sarosdy MF, Kahn PR, Ziffer MD, et al. Use of a multitarget fluorescence in situ hybridization assay to diagnose bladder cancer in patients with hematuria. J Urol. 2006 Jul; 176 (1): 44-47.

Jones S. DNA-based molecular cytology for bladder cancer surveillance. Urolog. 2006; 67 (Supplement 3A): 35-45.

Schlomer BJ, Ho R, Sagalowsky A, Ashfaq R, Lotan Y. Prospective validation of the clinical usefulness of reflex fluorescence in situ hybridization assay in patients with atypical cytology for the detection of urothelial carcinoma of the bladder. J Urol. 2010 Jan; 183 (1): 62-67.

Vysis Microscope Filter Recommendations

UroVysion probe signals and DAPI counterstain should be viewed with the following Vysis filter sets:

• DAPI single bandpass (DAPI counterstain)
• Aqua single bandpass (chromosome 17)
• Gold (Yellow) single bandpass (p16 gene)
• Red/Green dual bandpass (chromosomes 3 and 7)

An epi-fluorescence microscope equipped with a 100-watt mercury lamp is strongly recommended. An epi-fluorescence microscope enables a light source to transmit light through the viewing path, replacing the illuminator and condenser of the conventional wide field microscope. For this mode to function, the structures of interest are labeled with a fluorophore. Light from the source is collimated and directed into the tube of the microscope to a 45-degree mirror with an interference coating that lets in light at particular wavelengths and is transparent at other wavelengths (a dichroic mirror). This light is directed to the back of the objective and is focused into the sample. The objective functions as a condenser. Exciting light is absorbed by the fluorophore in the sample. The energy of the absorbed photon causes an electron in the dye molecule to jump to a higher energy orbital state. The electron rapidly jumps back to its ground orbital state, with the energy released becoming a photon of less energy compared to the exciting photon. The emitted photon has a color shifted toward red. The eyepiece collects the image in an identical manner as is transmitted in wide field microscopy.