METHOD FOR IDENTIFYING BIOLOGICAL MARKERS DETECTED IN HUMAN BIOLOGICAL MATERIALS IN VIEW OF POSSIBLE PRESENCE OF PATHOLOGICAL CONDITIONS OF THE HUMAN BODY, INCLUDING ONCOLOGICAL DISEASES, IMPLEMENTED BY MEANS OF MULTIPLEX IMMUNOENZYME SANDWICH IMMUNOASSAY Russian patent published in 2022 - IPC G01N33/543 

FIELD: medicine.

SUBSTANCE: invention relates to the field of medicine and biotechnology, namely, to comprehensive enzyme immunoassay in the microplate format. An array of immobilised monoclonal antibodies specifically binding oncological markers from the group AFP, CA15-3, CA19-9, CA72-4, CA-125, hCG, NSE, HE4, PSA, S100, Cyfra 21-1, CEA, SCC, immobilised in individual reaction zones of the immunosorbent, is brought into contact with a human serum or plasma sample. Said reaction zones are applied directly to the bottom of a flat-bottomed polystyrene reaction cell with a volume of 100 to 1,000 mcl, the diameter of each zone is 50 to 250 mcm, wherein the centres of the zones are distanced by 200 to 1,000 mcm by radius. The sample and said array of reaction zones are incubated, followed by washing the surface of the immunosorbent and introducing a solution of a mixture of biotinylated conjugates of monoclonal antibodies, incubating, washing the immunosorbent and introducing a solution of a conjugate of streptavidin with peroxidase, incubating for 5 to 20 minutes at 20 to 42°C in a stationary state or while stirring at a frequency of 5 to 30 Hz. The immunosorbent is washed, and a solution of a precipitating tetramethylbenzidine-based chromogene is introduced, followed by incubating and fully aspirating the liquid contents. The tumour markers bound to the immunosorbent are detected by measuring the intensity of generation of an optical signal in the immunosorbent zones with the supported monoclonal antibodies as a result of the formation of specific immune complexes as a result of an immunoenzyme reaction with peroxidase as part of the conjugate of the immune complex, leading to the precipitation of the 3,3’,5,5’-tetramethylbenzidinediimine chromogene and the formation of a stained section of the immunosorbent.

EFFECT: invention allows for multiparametric identification and evaluation of the concentration of biological marker analytes associated with oncological conditions of the human body, allows for a significant reduction in the time and labour consumption for estimating the content of tumour markers in the blood.

6 cl, 4 dwg, 5 tbl, 1 ex