DIAGNOSTIC TECHNIQUE FOR UROTHELIAL BLADDER CANCER Russian patent published in 2012 - IPC G01N33/50 

FIELD: medicine.

SUBSTANCE: invention describes a diagnostic technique for urothelial bladder cancer which is based on FISH-detection of genetic changes in urothelial cells specific for bladder cancer, namely increasing a number of pericentromeric signals of the 3rd, 7th and 17th chromosomes and/or the absence of 9p21 locus signal detected by UroVysion DNA probes. For this purpose, a morning second urine sample is sequentially processed with polyethylene glycol, phosphate buffer, Carnoy's fixative; each processing cycle is followed by centrifugation. A cell suspension is dropped into 10-mm wells of a slide to visualise 10-20 cells in a field of vision. Then the preparation is heated for 2-4 h at 39°C, added with 200 mcl of heated 0.005% RNAase, covered with a cover glass of the size equal to the slide, placed with the slide down in a humid chamber at temperature 37°C for 30 min. It is followed by processing with phosphate buffer, 10% pepsin, phosphate buffer, 1% formaldehyde and dehydratation with alcohol. Mixed UroVysion DNA probe 0.5 mcl and hybridisation buffer 1 mcl are introduced in an end well which is covered with the cover glass of the diametre of 10 mm, heated for 5 min at 74°C, and for 18-24 h at 39°C. Then the preparation is washed in detergent solutions; DAPI is added to the well which is analysed under fluorescence microscope. Min. 25 morphologically abnormal cells are to be measured, and if at least 4 of them have 2 pericentromeric signals in at least two chromosomes of three: 3rd, 7th and 17th, and/or the 9p21 locus signal is absent in 12 of 25 cells, urothelial bladder cancer is diagnosed.

EFFECT: invention provides higher sensitivity of the technique at the early stages as compared to the more advanced stages, availability for wide application.

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