METHOD FOR DIFFERENTIAL DIAGNOSIS OF INFLAMMATORY AND TUMOUR DISEASES Russian patent published in 2024 - IPC G01N33/52 G01N33/493 G01N33/533 G01N33/574 G01N33/577 G01N1/30 

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to oncology, urology, and can be used for differential diagnosis of inflammatory and malignant bladder diseases. Morning urine is sampled and flow cytometry is performed. First morning urine portion of patients with complaints is centrifuged at 2,000 rpm for 20 minutes to precipitate cell elements, then the supernatant is drained, 2 ml of PBS is added to the sediment, resuspended by pipetting for 1-2 minutes and centrifuged at 2,000 rpm for 10 minutes. Procedure for washing the cell sediment is carried out twice, then the supernatant is drained, 0.5 ml of PBS is added to the sediment, and the sediment is resuspended by pipetting for 1-2 minutes. Cell cycle is studied using a method of fluorescent staining with antibodies to ki-67 and propidium iodide. To the recovered cell elements from the urine sediment, 3 ml of 96% ethanol is added; the cell suspension is incubated in cold for 30 minutes and then washed twice in PBS. Method includes staining with monoclonal antibodies to ki-67, labelled with fluorescein isothiocyanate in amount of 20 mcl with addition of propidium iodide – phenanthridine-3,8-diamino-5-[3-(diethylmethylammonium) propyl]-6-phenyl-diiodide in concentration of 2 mg/ml in volume of 20 mcl, after which the suspension is stirred and incubated in dark for 15 minutes. After incubation, fluorescence is measured on a BD FACSCanto II flow cytometer. Presence of an inflammatory or malignant disease in the patients is determined from Table 2 of the description.

EFFECT: method enables the differential diagnosis of bladder diseases with high diagnostic accuracy, which enables the correct diagnosis at the first visit and the timely initiation of treatment, by assessing the distribution of cells in the urine sediment of the patients by different phases of the cell cycle using a flow cytometry method by fluorescent staining of cells with antibodies to ki-67 and propidium iodide.

1 cl, 2 tbl, 2 ex