METHOD FOR DETECTION OF UNSTABLE EXTRACELLULAR DNA, AND DEVICE USING IT Russian patent published in 2022 - IPC C12Q1/6837 

FIELD: biotechnology.

SUBSTANCE: method for the detection of extracellular DNA (hereinafter – ecDNA) having an unstable two-spiral structure from a sample without amplification is described, including: mixing of a sample containing ecDNA with positively charged substance; isolation of positively charged substance, to which ecDNA is related; mixing of the mixture with a probe and a marker; removal of the probe and the marker unrelated to ecDNA; and detection of the marker. A method for obtaining information for diagnostics of malignant tumor by detection of ecDNA having an unstable two-spiral structure from a sample without amplification is also described, including: mixing of a sample containing ecDNA with positively charged substance; isolation of positively charged substance, to which ecDNA is related; mixing of the mixture with a probe and a marker; removal of the probe and the marker unrelated to ecDNA; detection of the marker; and determination of that there is malignant tumor associated with a gene corresponding to ecDNA having an unstable two-spiral structure, when the marker is detected. In addition, a method for obtaining information for diagnostics of an infectious disease by detection of ecDNA having an unstable two-spiral structure from a sample without amplification is described, including: mixing of a sample containing ecDNA with positively charged substance; isolation of positively charged substance, to which ecDNA is related; mixing of the mixture with a probe and a marker; removal of the probe and the marker unrelated to the ecDNA; detection of the marker; and determination of that there is an infectious disease associated with a gene corresponding to ecDNA having an unstable two-spiral structure, when the marker is detected.

EFFECT: invention makes it possible to detect small-sized ecDNA with ultra-high sensitivity from a liquid sample, such as urine, cerebrospinal fluid, plasma, blood, pleural fluid, or body fluid, it is concentrated and isolated, and then analyzed for mutations in genes without PCR; in particular, in case, when a positively charged nanostructure is used, binding and detection rates for ecDNA can be increased; at the same time, no PCR amplification reaction is required, which greatly shortens the time spent to obtain the result; in addition, since direct on-site analysis is possible without the need for specific equipment, it is expected that the present invention can be used as on-site testing (ROST) capable of simultaneously searching for a set of genes in a short period of time.

21 cl, 61 dwg, 4 tbl, 14 ex