METHOD OF DETECTION OF CELL-FREE DNA IN WHOLE PERIPHERAL BLOOD USING FLOW CYTOMETRY Russian patent published in 2024 - IPC G01N33/50 G01N33/577 

FIELD: laboratory diagnostics.

SUBSTANCE: method of detecting extracellular DNA in whole peripheral blood using flow cytometry is described. The method involves preparing a solution of propidium iodide dye with a concentration of 100 mcg/ml containing 100 mcg/ml RNase, simultaneous addition of 10 mcl of a solution of propidium iodide and 10 FITC-labelled monoclonal antibodies to the common leukocyte antigen CD45-FITC and/or 10 mcl of phycoerythrin-labelled monoclonal antibodies to the surface phenotypic marker of early apoptosis CD95-PE to 50 mcl of heparinized whole blood, mixing and incubating the sample in the dark at room temperature for 15 minutes. Next is the addition of 450 mcl of BD FACS Lysing Solution (BD Cat. No. 343236) diluted 10 times with deionized water to the sample, mixing and incubation at room temperature in the dark for 30 minutes, followed by cytometric DNA analysis using the Lyse/No Wash protocol.

EFFECT: provision of fast and accurate detection of extracellular DNA in whole peripheral blood, as well as simplification of the technology of preparing a sample for research.

1 cl, 3 dwg, 1 tbl, 3 ex