Bladder Cancer

Bladder cancer is the fifth most common cancer in the United States, with over 50,000 newly diagnosed cases and over 12,000 deaths annually . The recurrence rate for bladder cancer is 70 percent; of the approximately 600,000 Americans living with bladder cancer, it is estimated that nearly 400,000 are at risk for recurrence.

In men, bladder cancer is the fifth most common malignant tumor in Europe and the fourth most common in the USA. In American women it is the 8th most common cancer.

There are many different tests available for the diagnosis of bladder cancer. The UroVysion^TM bladder cancer recurrance kit is the first and only DNA probe test to directly examine bladder cell DNA for signs of cancer.

The UroVysion^TM Bladder Cancer Kit (Vysis-Abbott) 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^TM 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.

Automated FISH (A-FISH):

BioView offers automated applications for the analysis of cells in voided urine samples for diagnosis and monitoring of subjects with transitional cell carcinoma of the bladder. The Duet ability to efficiently scan, classify and analyze cells hybridized with the Vysis UroVysion^TM Bladder Cancer Kit, enables fast and accurate detection of aneuploidy for chromosomes 3, 7, 17 and loss of the 9p21 locus via fluorescence in situ hybridization (FISH). Coupled with the Solo – BioView’s offline analysis workstation – the Duet increases the throughput of the laboratory and reduces its operational costs.

Abnormal Normal

Advantages of Automated Scanning

Rapid scan and analysis of cells hybridized with fluorescent markers
Use BioView SmartStop to determine whether to scan the entire slide, to stop after a pre-set number of cells or stop after a pre-set number of abnormal cells
Quickly review and approve the scan results using the Duet™ /Solo™ user-friendly screens
View two images of the same cell taken with different filter sets side-by-side
View all images of the same cell taken with different focal planes (z-stack)
View a combined image showing all the signals in the cell

A screen of voided urine cells presenting abnormal chromosomal status after labeling with UroVysion probe. The left image of each pair shows the gold filter and the right image shows the combined image of all the filters and all the focus plains images, of the same cell.

Duet system has FDA clearance for the Automated FISH (A-FISH) Bladder Cancer Application

Indication for use: The Duet^TM system is an automated scanning microscope and image analysis system. It is intended for in-vitro diagnostic use as an aiding tool to the pathologist in the detection, classification and counting of cells of interest based on color, intensity, size, pattern and shape. This particular application is intended for detecting cells in urine specimens, stained by FISH (using the Vysis UroVysion^TMBladder Cancer Recurrence Kit for chromosomes 3,7,17 and 9p21 locus) from subjects with transitional cell carcinoma of the bladder.

The Urovysion Kit is from Vysis Inc., Downer’s Grove, IL

Target FISH (T-FISH)

FOR RESEARCH USE ONLY

It has already been reported that cells with FISH abnormalities (using the UroVysion probe panel or other FISH probes) generally exhibit nuclear abnormalities including nuclear enlargement, irregular nuclear borders and patchy DAPI staining. Therefore, the most efficient way to evaluate these FISH slides is to scan for cells with nuclear abnormalities. However, using FISH staining in order to find morphologic abnormalities in the nuclei is usually limited.

BioView now offers a new dimension in the analysis of bladder cancer samples: viewing cell’s abnormalities under cytology bright-filed staining and returning exactly to the same cell for FISH analysis.

The next set of images will exemplify the advantages of our approach. In each pair of images, the morphology image of the cell is presented on the right side and the FISH image of the same cell is presented on the left.

The first pair demonstrates an abnormal cell both in morphology and FISH. In this particular case, an experienced lab technician would suspect this cell to be malignant under DAPI counterstain. The nucleus is irregular and enlarged, and the DAPI counterstain is patchy. The morphology image confirms the suspicion.

Example 1

Abnormal DAPI and Abnormal FISH Abnormal morphology

The next pair demonstrates a cell with normal FISH signals, however it looks suspicious under DAPI conterstain because the nucleus exhibited enlargement and irregular borders. When combined with the morphology findings, it is clear that this cell is normal.

Example 2

Abnormal DAPI and Normal FISH Normal morphology

Below is an example of a cell that does not look suspicious under DAPI counterstain but the morphology is abnormal. When looking at the FISH signals it is obvious that this cell is abnormal.

Example 3

Normal DAPI and Abnormal FISH Abnormal mprphology

Finally, the last example demonstrates the ability of the method to accurately identify rare cells. It shows a cell, unquestionably abnormal both under morphology and DAPI staining. This was the only abnormal cell found in this sample. All other cells were morphology normal and had a normal FISH pattern. If this slide was scanned only for FISH, this cell may have been ignored. But taking together the highly abnormal morphology with the polyploidy FISH signal pattern provides much stronger evidence for the possible existence of a tumor in this patient.

Example 4

Abnormal DAPI and Abnormal FISH Abnormal morphology

The combined analysis enables resolution of those cases where only a few abnormal cells are found. Since there are two indications on the same cell, this increases the specificity of the result.

In conclusion, the results of our preliminary studies have demonstrated that the combination of morphology and FISH is powerful tool providing high sensitivity and specificity in the detection of bladder cancer. The advantages of this approach is especially noticeable in superficial, low-stage, law-grade tumors. Thhe straightforward preparation protocol and the uniqe features of the Duet system create a particular test that is easily applicable in any routine cytology laboratory.

Urology 66, 1354-1359