METHOD FOR EARLY DETECTION OF BLADDER CANCER USING CYTOFLUORIMETRIC ANALYSIS OF THE URINARY CELLULAR SEDIMENT Russian patent published in 2021 - IPC G01N33/50 

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology, urology, and can be used for early detection (screening) of bladder cancer, and applies to a method for early detection of bladder cancer using cytofluorimetric analysis of urinary cellular sediment. The method includes: collecting the first morning portion of the urine of the subject, centrifuging at 2,000 rpm for 20 minutes for sedimentation of cellular elements; draining the supernatant, adding 2 ml PBS to the sediment, resuspending by pipetting for 1 to 2 minutes and re-centrifuging at 2,000 rpm for 10 minutes; the washing procedure is executed twice; draining the supernatant, adding 0.5 ml PBS to the sediment, resuspending by pipetting for 1 to 2 minutes. Monoclonal antibodies to CD45 labeled with PerCP-Cyanine5.5 are then added to the resulting cellular suspension in a volume of 20 mcl, mixed on a personal vortex for 20 seconds, monoclonal antibodies to CD326 labeled with allophycocyanin are added in a volume of 20 mcl, mixed on a personal vortex for 20 seconds and placed in a dark place; incubation of the cellular suspension is executed at room temperature for 15 minutes; after incubation, fluorescence is measured on a flow cytofluorometer BD FACSCanto II, the obtained data is analysed using the FACSDiva software by light scattering, the amount of CD45⁺ events and the amount of CD326⁺ events; if the amount of CD45^-CD326⁺ cells is less than or equal to 2.65%, the presence of a malignant neoplasm of the bladder at an early stage without clinical manifestations is not assumed; if the number of CD45^-CD326⁺ cells is above a 2.65%, the presence of malignant neoplasms of the bladder at an early stage without clinical manifestations in the studied subject is assumed. The invention is an effective, accurate, easy to implement, non-invasive detection of asymptomatic malignant neoplasms of the bladder based on cytometric analysis of urinary sediment using CD45 and CD326 monoclonal antibodies.

EFFECT: provides a possibility to detect the presence of bladder cancer without clinical manifestations in the early stages with high diagnostic accuracy (98.1%).

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